Substituted bicyclic compounds

ABSTRACT

Disclosed are compounds of Formulas (I), (II), (III), (IV), and (V): 
                         
and/or a salt thereof, wherein R 1  is —OH or —OP(O)(OH) 2 , and X 1 , X 2 , X 3 , R 2 , R 2a , R a , R b , and R c  are defined herein. Also disclosed are methods of using such compounds as selective agonists for G protein-coupled receptor S1P 1 , and pharmaceutical compositions comprising such compounds. These compounds are useful in treating, preventing, or slowing the progression of diseases or disorders in a variety of therapeutic areas, such as autoimmune diseases and vascular disease.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. application Ser.No. 16/879,824, filed May 21, 2020, which is a continuation applicationof U.S. application Ser. No. 16/188,444, filed Nov. 13, 2018, which is acontinuation application of U.S. application Ser. No. 15/648,250, filedJul. 12, 2017, which is a continuation application of U.S. applicationSer. No. 15/337,843, filed Oct. 28, 2016, which is a divisionalapplication of U.S. application Ser. No. 14/831,439, filed Aug. 20,2015, which claims the benefit of U.S. Application Ser. No. 62/039,622,filed Aug. 20, 2014, each of which is incorporated herein it itsentirety

DESCRIPTION

The present invention generally relates to substituted bicycliccompounds useful as S1P₁ agonists. Provided herein are substitutedbicyclic compounds, compositions comprising such compounds, and methodsof their use. The invention further pertains to pharmaceuticalcompositions comprising at least one compound according to the inventionthat are useful for the treatment of conditions related to S1P₁modulation, such as autoimmune diseases and vascular disease.

Sphingosine-1-phosphate (S1P) is a zwitterionic lysophospholipidmetabolite of sphingosine (Sph), which in turn is derived from enzymaticcleavage of ceramides. Enzymatic phosphorylation of Sph by two kinases(SphK1 and SphK2) leads to the production of S1P largely fromerythrocytes, but also from a radiation resistant source, possibly thelymphatic endothelium (Pappu, R. et al. Science 2007, 316, 295-298).Originally thought to operate solely as an intracellular signalingmolecule, S1P was subsequently identified as a high affinity ligand forfive members of the endothelial differentiation gene (EDG) class ofG-protein coupled receptors (GPCRs) named S1P₁ or S1P1, S1P₂ or S1P2,S1P₃ or S1P3, S1P₄ or S1P4, and S1P₅ or S1P5 (formerly called EDG-1,EDG-5, EDG-3, EDG-6, and EDG-8, respectively) (Chun, J. et al.Pharmacological Rev. 2010, 62, 579-587). The interaction of S1P with theS1P receptors plays a fundamental physiological role in a large numberof processes including cell proliferation, cell morphology, tumor cellinvasion, angiogenesis, tumorigenesis, cytoskeletal rearrangement,vascular development, and lymphocyte trafficking (Olivera, A; Rivera, J.Adv Exp Med Biol. 2011, 716, 123-142). S1P receptors are therefore goodtargets for a wide variety of therapeutic applications such as tumorgrowth inhibition, vascular disease, and autoimmune diseases.

Among the five S1P receptors, S1P₁ has a widespread distribution. It isthe predominant family member expressed on lymphocytes and plays animportant role in lymphocyte trafficking. S1P interaction with itsreceptor S1P₁ is required for the egress of immune cells from thelymphoid organs (such as thymus and lymph nodes) into the lymphaticvessels. Downregulation of the S1P₁ receptor (which can be accomplishedthrough treatment with agonists of S1P₁ via receptor internalization)disrupts lymphocyte migration and homing to various tissues. Thisresults in sequestration of the lymphocytes in lymph organs therebydecreasing the number of circulating lymphocytes that are capable ofmigration to the affected tissues. The development of an S1P₁ receptormodulating agent that suppresses lymphocyte migration to the targetsites associated with autoimmune and aberrant inflammatory processescould be efficacious in a number of autoimmune and inflammatory diseasestates.

The following applications have described compounds as S1P₁ agonists: WO03/061567 (U.S. Patent Publication No. 2005/0070506), WO 03/062248 (U.S.Pat. No. 7,351,725), WO 03/062252 (U.S. Pat. No. 7,479,504), WO03/073986 (U.S. Pat. No. 7,309,721), WO 03/105771, WO 05/058848, WO05/000833, WO 05/082089 (U.S. Patent Publication No. 2007/0203100), WO06/047195, WO 06/100633, WO 06/115188, WO 06/131336, WO 2007/024922, WO07/109330, WO 07/116866, WO 08/023783 (U.S. Patent Publication No.2008/0200535), WO 08/029370, WO 08/074820, WO 08/079382, WO 08/114157,WO 09/043889, WO 09/057079, and U.S. Pat. No. 6,069,143. Also see Haleet al., J. Med. Chem., 47:6662 (2004).

There still remains a need for compounds useful as S1P₁ agonists and yethaving selectivity over S1P3.

SUMMARY OF THE INVENTION

The present invention provides substituted bicyclic compounds, which areuseful as modulators of S1P₁ activity, including salts thereof.

The present invention also provides pharmaceutical compositionscomprising a compound of Formulas (I), (II), (III), (IV), or (V) and/ora pharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable carrier.

The present invention also provides a method of treating a disease ordisorder associated with the activity of G protein-coupled receptorS1P₁, the method comprising administering to a mammalian patient acompound of Formulas (I), (II), (III), (IV), or (V) and/or apharmaceutically acceptable salt thereof.

The present invention also provides processes and intermediates formaking the compounds of Formulas (I), (II), (III), (IV), or (V) and/orsalts thereof.

The present invention also provides a compound of Formulas (I), (II),(III), (IV), or (V) and/or a pharmaceutically acceptable salt thereof,for use in therapy.

The present invention also provides the use of the compounds of Formulas(I), (II), (III), (IV), or (V) and/or pharmaceutically acceptable saltsthereof, for the manufacture of a medicament for the treatment orprophylaxis of S1P₁ receptor-related conditions, such as autoimmune andvascular diseases.

The compounds of Formulas (I), (II), (III), (IV), or (V) andcompositions comprising the compounds of Formulas (I), (II), (III),(IV), or (V) may be used in treating, preventing, or curing various S1P₁related conditions. Pharmaceutical compositions comprising thesecompounds are useful in treating, preventing, or slowing the progressionof diseases or disorders in a variety of therapeutic areas, such asautoimmune and vascular diseases.

These and other features of the invention will be set forth in expandedform as the disclosure continues.

DETAILED DESCRIPTION

The first aspect of the present invention provides at least one compoundof Formulas (I), (II), (III), (IV), or (V):

-   or a salt thereof, wherein:-   R₁ is —OH or —OP(O)(OH)₂;-   X₁ is CH₂ or O;-   X₂ is CH₂ or O;-   X₃ is CH₂ or O, provided that X₂ is O only if both X₁ and X₃ are    each CH₂;-   R₂ is R_(2a) or R_(2b);-   represents either a single bond to R_(2a) or a double bond to    R_(2b);-   R_(2a)    -   is —(CH₂)₃₋₆CH₃, —(CH₂)₁₋₄CH═CR_(x)R_(x),        —(CH₂)₁₋₄CH═CR_(x)(CH₂CH₃), —CH═CH(CH₂)₁₋₃C(R_(x))₃,        —CH═CH(CH₂)₁₋₃OCH₃, —(CH₂)₁₋₃CH═CHCH═CR_(x)R_(x),        —CH═CH(CH₂)₁₋₃CH═CR_(x)R_(x), —CH═CHR_(z), —(CH₂)₁₋₃R_(z),        —(CH₂)₁₋₃O(CH₂)₀₋₃R_(z), —(CH₂)₁₋₃S(CH₂)₀₋₃R_(z), —CH₂S(O)R_(z),        —CH₂S(O)₂R_(z), —O(CH₂)₁₋₂R_(z), —O(CH₂)₁₋₂O(CH₂)₀₋₂R_(z),        —OC(O)R_(z), —(CH₂)₁₋₄O(CH₂)₀₋₉C(R_(x))₃, —(CH₂)₁₋₄O(CH₂)₀₋₉CF₃,        —(CH₂)₁₋₄CR_(x)R_(x)O(CH₂)₀₋₄C(R_(x))₃,        —(CH₂)₁₋₃O(CH₂)₁₋₄CH═CR_(x)(CH₂)₀₋₃CH₃,        —(CH₂)₁₋₃O(CH₂)₁₋₄CH═CR_(x)R_(x),        —(CH₂)₁₋₃O(CH₂)₁₋₄C(OH)R_(x)R_(x),        —(CH₂)₁₋₃O(CH₂)₁₋₄O(CH₂)₀₋₃CH₃, —(CH₂)₁₋₃S(CH₂)₀₋₄C(R_(x))₃,        —(CH₂)₀₋₃O(CH₂)₁₋₄S(CH₂)₀₋₃C(R_(x))₃,        —(CH₂)₁₋₃S(CH₂)₁₋₄Si(CH₃)₃, —(CH₂)₁₋₃S(O)(CH₂)₀₋₄C(R_(x))₃,        —(CH₂)₁₋₃S(O)₂(CH₂)₀₋₄C(R_(x))₃, —(CH₂)₁₋₅NR_(x)R_(x),        —O(CH₂)₁₋₇C(R_(x))₃, —O(CH₂)₁₋₄O(CH₂)₀₋₄C(R_(x))₃,        —O(CH₂)₁₋₄CH═CR_(x)(CH₂)₀₋₃CH₃, —O(CH₂)₁₋₄O(CH₂)₀₋₃C(R_(x))₃,        —O(CH₂)₁₋₄O(CH₂)₁₋₃CH═CR_(x)R_(x), —O(CH₂)₁₋₄O(CH₂)₁₋₃C≡CR_(x),        —C(O)(CH₂)₀₋₄C(R_(x))₃, —OC(O)(CH₂)₀₋₄C(R_(x))₃,        —OC(O)CR_(x)R_(x)(CH₂)₀₋₄C(R_(x))₃,        —OC(O)NR_(x)(CH₂)₀₋₅C(R_(x))₃,        —NR_(x)C(O)NR_(x)(CH₂)₀₋₅C(R_(x))₃,        —C(CH₃)═N—O(CH₂)₀₋₅C(R_(x))₃, —C(CH₃)═N—O(CH₂)₁₋₂(phenyl),        —C(CH₃)═N—O(CH₂)₁₋₂(fluorophenyl),        —C(CH₃)═N—O(CH₂)₁₋₂(methoxyphenyl), phenyl, or pyridinyl;-   R_(2b) is    -   (i) a 6-membered spiro-ring having one oxygen atom and        substituted with zero or 1 substituent selected from —(CH₂)₃CH₃;        or    -   (ii)═N—O—(CH₂)₃CH₃, ═N—O—CH₂CH(CH₃)₂, ═N—OCH₂CH₂(phenyl), or        ═N—O—CH₂CH₂CH₂(phenyl);-   R_(a) is H or —OH;-   each R_(b) is independently H or —CH₃;-   each R_(c) is independently H, Cl, I, or —CH₃;-   each R_(x) is independently H or —CH₃; and-   R_(z) is phenyl, imidazolyl, pyrazolyl, pyridinyl, pyrimidinyl,    pyrazinyl, quinolinyl, thiophenyl, thiazolyl, oxetanyl, C₃₋₆    cycloalkyl, adamantanyl, or tetrahydropyranyl, each substituted with    zero to 4 substituents independently selected from F, Cl, I, C₁₋₄    alkyl, —O(C₁₋₃ alkyl), —CF₃, —OCF₃, —(CH₂)₁₋₆OCH₃, —CH₂NR_(x)R_(x),    —C(O)NR_(x)R_(x), —C(O)NR_(x)(C₁₋₄ alkyl), and —CH₂C(O)NR_(x)R_(x);    with the provisos that (i) if said compound has the structure of    Formula (I) and R₂ is —(CH₂)₅CH₃, then at least one of R_(b) and R,    is not H; and (ii) if said compound has the structure of    Formula (II) and X₁, X₂, and X₃ are each CH₂, then R_(2a) is not    —(CH₂)₅CH₃.

One embodiment provides at least compound of Formulas (I), (II), (III),(IV), or (V) or a salt thereof, wherein R₁ is —OH, and R₂, R_(2a), X₁,X₂ X₃, R_(a), R_(b), and R_(c) are defined in the first aspect. Thecompounds of this embodiment have structures of Formulas (Ia), (IIa),(IIIa), (IVa), or (Va):

One embodiment provides at least compound of Formulas (I), (II), (III),(IV), or (V) or a salt thereof, wherein R₁ is —OP(O)(OH)₂, and R₂,R_(2a), X₁, X₂ X₃, R_(a), R_(b), and R_(c) are defined in the firstaspect. The compounds of this embodiment have structures of Formulas(Ib), (IIb), (IIIb), (IVb), or (Vb):

The compounds of Formulas (Ia), (IIa), (IIIa), (IVa), and (Va) or saltsthereof are useful as prodrugs of the corresponding compounds ofFormulas (Ib), (IIb), (IIIb), (IVb), and (Vb), or salts thereof. Thecompounds of Formula (Ia), (IIa), (IIIa), (IVa), and (Va) are activatedin vivo through phosphorylation to provide the correspondingphosphorylated compounds. The phosphorylated compounds of Formula (Ib),(IIb), (IIIb), (IVb), and (Vb) or salts thereof are active as selectiveagonists of S1P₁.

One embodiment provides at least one compound of Formulas (I), (II),(III), (IV), or (V), wherein said compound has the structure of Formula(Ic), (IIc), (IIIc), (IVc), or (Vc):

One embodiment provides at least one compound of compound of Formulas(I), (II), (III), (IV), or (V), or a salt thereof, wherein R_(2a)

is —(CH₂)₃CH₃, —(CH₂)₅₋₆CH₃, —CH₂CH═CHCH₂CH₃, —CH₂CH₂CH═CHCH₂CH₃,—(CH₂)₃CH═CHCH₃, —(CH₂)₃CH═C(CH₃)₂, —(CH₂)₄CH═CH₂, —(CH₂)₄CH═CHCH₃,—CH═CH(CH₂)₃CH₃, —CH═CH(CH₂)₃OCH₃, —CH═CHCH₂CH₂CH(CH₃)₂,—CH═CHCH₂CH₂CH₂OCH₃, —CH₂CH═CHCH═CHCH₃, —CH═CHCH₂CH₂CH═CH₂,—CH═CH(phenyl) wherein said phenyl is substituted with —CH₃or —OCH₃; —CH═CH(tetrahydropyranyl), —(CH₂)₁₋₃(phenyl) wherein saidphenyl is substituted with zero to 2 substituents independently selectedfrom F, I, —CH₃, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, and —CH₂C(O)N(CH₃)₂;—(CH₂)₂(methyl imidazolyl), —(CH₂)₂(methyl pyrazolyl),—(CH₂)₁₋₂(pyridinyl) wherein said pyridinyl is substituted with zero to1 substituent selectedfrom —OCH₃; —(CH₂)₂(pyrimidinyl), —(CH₂)₂(quinolinyl),—(CH₂)₂₋₃(tetrahydropyranyl), —CH₂O(CH₂)₃₋₄CH₃, —CH₂OCH₂CH₂CH(CH₃)₂,—CH₂OCH₂CH₂C(CH₃)₃, —CH₂O(CH₂)₉CH₃, —CH₂OCH₂CH₂CH₂CF₃,—CH₂OCH₂CH═CHCH₂CH₃, —CH₂OCH₂CH═C(CH₃)₂, —CH₂OCH₂CH═CHCH₂CH₂CH₃,—CH₂OCH₂CH₂CH═CH₂, —CH₂OCH₂CH₂CH₂CH═CH₂, —CH₂OCH₂CH₂CH═C(CH₃)₂,—CH₂OCH₂CH₂CH(OH)CH₃, —CH₂OCH₂CH₂CH₂CH₂OH, —CH₂OCH₂CH₂CH₂C(CH₃)₂(OH),—CH₂OCH₂CH₂OCH₃, —CH₂OCH₂CH₂CH₂OCH₃, —CH₂OCH₂CH₂OCH₂CH₂CH₃,—CH₂O(phenyl) wherein said phenyl is substituted with zero to 3substituents independently selected from F, Cl, —CH₃, —CH(CH₃)₂,—C(CH₃)₃, —OCH₃, —OCF₃, —(CH₂)₁₋₆OCH₃, —C(O)N(CH₃)₂, —CH₂N(CH₃)₂,—C(O)N(CH₂CH₃)(CH₃), —C(O)N(CH₃)(CH₂CH₂CH₂CH₃), and—C(O)N(CH₃)(CH₂CH(CH₃)₂); —CH₂O(methoxy pyridinyl),—CH₂O(tetrahydropyranyl), —CH₂O(trifluoromethyl, methyl pyrazolyl),—CH₂OCH₂(phenyl) wherein said phenyl is substituted with zero to 1substituent selected from —CH₃ and —OCH₃; —CH₂OCH₂(methyl pyrazolyl),—CH₂OCH₂(tetrahydropyranyl), —CH₂OCH₂(thiophenyl), —CH₂OCH₂(trifluoromethyl thiophenyl), —CH₂OCH₂(ethyl thiophenyl), —CH₂OCH₂(dimethylthiophenyl), —CH₂CH₂OCH₂CH₃, —CH₂CH₂OCH₂CH(CH₃)₂,—CH₂CH₂O(methoxyphenyl), —CH₂CH₂OCH₂(cyclopropyl), —CH₂CH₂SCH(CH₃)₂,—(CH₂)₃OCH₂CH₃, —(CH₂)₃OCH(CH₃)₂, —(CH₂)₃OCH₂CH₂CH═CH₂,—(CH₂)₃O(oxetanyl), —(CH₂)₃O(tetramethyl cyclohexyl), —(CH₂)₃OCH₂SCH₃,—CH₂S(CH₂)₂₋₄CH₃, —CH₂SCH(CH₃)₂, —CH₂SCH₂CH(CH₃)₂, —CH₂SCH₂C(CH₃)₃,—CH₂SCH₂CH₂CH(CH₃)₂, —CH₂SCH₂CH₂C(CH₃)₃, —CH₂SCH₂CH₂Si(CH₃)₃,—CH₂CH₂S(CH₂)₁₋₂CH₃, —CH₂CH₂SCH₂CH(CH₃)₂, —CH₂S(phenyl) wherein saidphenyl is substituted with zero to 2 substituents independently selectedfrom —CH₃, —CH(CH₃)₂,and —OCH₃; —CH₂S(adamantanyl), —CH₂S(pyridinyl), —CH₂S(methylpyridinyl), —CH₂SCH₂CH₂(phenyl), —CH₂SCH₂CH₂(pyrazinyl),—CH₂SCH₂CH₂(pyridinyl), —CH₂S(O)(CH₂)₃CH₃, —CH₂S(O)₂(CH₂)₃CH₃,—CH₂S(O)(phenyl), —CH₂S(O)₂(phenyl), —(CH₂)₄OCH(CH₃)₂,—(CH₂)₄CH(CH₃)OCH₃, —(CH₂)₄C(CH₃)₂OCH₃, —(CH₂)₅N(CH₃)₂, —O(CH₂)₄₋₇CH₃,—OCH₂CH₂O(CH₂)₂₋₄CH₃, —OCH₂CH₂OCH₂CH(CH₃)₂, —OCH₂CH═CH(CH₂)₂₋₃CH₃,—OCH₂CH₂OCH₂CH═CH₂, —OCH₂CH₂OCH₂CH═CH(CH₃), —OCH₂CH₂OCH₂CH═C(CH₃)₂,—OCH₂CH₂OCH₂CH₂C≡CH, —OCH₂CH₂O(CH₂)₂₋₃CH(CH₃)₂, —OCH₂CH₂S(CH₂)₂CH₃,—OCH₂(cyclohexyl), —OCH₂(tetrahydropyranyl), —OCH₂(phenyl) wherein saidphenyl is substituted with zero to 1 substituent selectedfrom —CH₃, —CH₂CH₃, —OCH₃, —OCF₃,and —OCH₂CH₃; —OCH₂CH₂O(cyclohexyl), —OCH₂CH₂O(methyl phenyl),—OCH₂CH₂OCH₂(cyclobutyl), —OCH₂CH₂OCH₂(phenyl), —OCH₂CH₂OCH₂(thiazolyl),—OCH₂CH₂OCH₂(thiophenyl), —C(O)(CH₂)₄CH₃, —OC(O)(CH₂)₄CH₃,—OC(O)C(CH₃)₂(CH₂)₃CH₃, —OC(O)(phenyl), —OC(O)NH(CH₂)₃CH₃,—OC(O)NH(CH₂)₅CH₃, —OC(O)N(CH₃)(CH₂)₃CH₃, —OC(O)N(CH₃)(CH₂)₄CH₃,—NHC(O)NH(CH₂)₃CH₃, —C(CH₃)═N—O(CH₂)₃CH₃, —C(CH₃)═N—OCH₂(phenyl),—C(CH₃)═N—OCH₂(fluorophenyl), —C(CH₃)═N—OCH₂(methoxyphenyl),—C(CH₃)═N—OCH₂CH₂(phenyl), —OC(O)NH(CH₂)₃CH₃, —OC(O)NH(CH₂)₅CH₃,—OC(O)N(CH₃)(CH₂)₃₋₄CH₃, —NHC(O)NH(CH₂)₃CH₃, phenyl, or pyridinyl; andR_(2b) is

-   -   (i) a 6-membered spiro-ring having one oxygen atom and        substituted with zero or 1 substituent selected from —(CH₂)₃CH₃;        or    -   (ii)═N—O—(CH₂)₃CH₃, ═N—O—CH₂CH(CH₃)₂, ═N—OCH₂CH₂(phenyl), or        ═N—O—CH₂CH₂CH₂(phenyl);        and R₁, X₁, X₂, X₃, R_(a), R_(b), and R, are defined in the        first aspect. Included in this embodiment are compounds of        Formula (Ic), (IIc), (IIIc), (IVc), or (Vc).

One embodiment provides at least one compound of Formula (I) or a saltthereof, wherein: R₁ is —OH or —OP(O)(OH)₂; R₂ is R_(2a) or R_(2b);R_(2a)

is —(CH₂)₃CH₃, —(CH₂)₅CH₃, —CH₂CH═CHCH₂CH₃, —CH₂CH₂CH═CHCH₂CH₃,—(CH₂)₃CH═CHCH₃, —(CH₂)₃CH═C(CH₃)₂, —(CH₂)₄CH═CH₂, —(CH₂)₄CH═CHCH₃,—CH═CH(CH₂)₃CH₃, —CH═CH(CH₂)₃OCH₃, —CH═CHCH₂CH₂CH(CH₃)₂,—CH═CHCH₂CH₂CH₂OCH₃, —CH₂CH═CHCH═CHCH₃, —CH═CHCH₂CH₂CH═CH₂,—CH═CH(phenyl) wherein said phenyl is substituted with —CH₃or —OCH₃; —CH═CH(tetrahydropyranyl), —(CH₂)₁₋₃(phenyl) wherein saidphenyl is substituted with zero to 2 substituents independently selectedfrom F, I, —CH₃, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, and —CH₂C(O)N(CH₃)₂;—(CH₂)₂(methyl imidazolyl), —(CH₂)₂(methyl pyrazolyl),—(CH₂)₁₋₂(pyridinyl) wherein said pyridinyl is substituted with zero to1 substituent selectedfrom —OCH₃; —(CH₂)₂(pyrimidinyl), —(CH₂)₂(quinolinyl),—(CH₂)₂₋₃(tetrahydropyranyl), —CH₂O(CH₂)₃₋₄CH₃, —CH₂OCH₂CH₂CH(CH₃)₂,—CH₂OCH₂CH₂C(CH₃)₃, —CH₂O(CH₂)₉CH₃, —CH₂OCH₂CH₂CH₂CF₃,—CH₂OCH₂CH═CHCH₂CH₃, —CH₂OCH₂CH═C(CH₃)₂, —CH₂OCH₂CH═CHCH₂CH₂CH₃,—CH₂OCH₂CH₂CH═CH₂, —CH₂OCH₂CH₂CH₂CH═CH₂, —CH₂OCH₂CH₂CH═C(CH₃)₂,—CH₂OCH₂CH₂CH(OH)CH₃, —CH₂OCH₂CH₂CH₂CH₂OH, —CH₂OCH₂CH₂CH₂C(CH₃)₂(OH),—CH₂OCH₂CH₂OCH₃, —CH₂OCH₂CH₂CH₂OCH₃, —CH₂OCH₂CH₂OCH₂CH₂CH₃,—CH₂O(phenyl) wherein said phenyl is substituted with zero to 3substituents independently selected from F, Cl, —CH₃, —CH(CH₃)₂,—C(CH₃)₃, —OCH₃, —OCF₃, —(CH₂)₁₋₆OCH₃, —C(O)N(CH₃)₂, —CH₂N(CH₃)₂,—C(O)N(CH₂CH₃)(CH₃), —C(O)N(CH₃)(CH₂CH₂CH₂CH₃), and—C(O)N(CH₃)(CH₂CH(CH₃)₂); —CH₂O(methoxy pyridinyl),—CH₂O(tetrahydropyranyl), —CH₂O(trifluoromethyl, methyl pyrazolyl),—CH₂OCH₂(phenyl) wherein said phenyl is substituted with zero to 1substituent selected from —CH₃ and —OCH₃; —CH₂OCH₂(methyl pyrazolyl),—CH₂OCH₂(tetrahydropyranyl), —CH₂OCH₂(thiophenyl), —CH₂OCH₂(trifluoromethyl thiophenyl), —CH₂OCH₂(ethyl thiophenyl), —CH₂OCH₂(dimethylthiophenyl), —CH₂CH₂OCH₂CH₃, —CH₂CH₂OCH₂CH(CH₃)₂,—CH₂CH₂O(methoxyphenyl), —CH₂CH₂OCH₂(cyclopropyl), —CH₂CH₂SCH(CH₃)₂,—(CH₂)₃OCH₂CH₃, —(CH₂)₃OCH(CH₃)₂, —(CH₂)₃OCH₂CH₂CH═CH₂,—(CH₂)₃O(oxetanyl), —(CH₂)₃O(tetramethyl cyclohexyl), —(CH₂)₃OCH₂SCH₃,—CH₂S(CH₂)₂₋₄CH₃, —CH₂SCH(CH₃)₂, —CH₂SCH₂CH(CH₃)₂, —CH₂SCH₂C(CH₃)₃,—CH₂SCH₂CH₂CH(CH₃)₂, —CH₂SCH₂CH₂C(CH₃)₃, —CH₂SCH₂CH₂Si(CH₃)₃,—CH₂CH₂S(CH₂)₁₋₂CH₃, —CH₂CH₂SCH₂CH(CH₃)₂, —CH₂S(phenyl) wherein saidphenyl is substituted with zero to 2 substituents independently selectedfrom —CH₃, —CH(CH₃)₂,and —OCH₃; —CH₂S(adamantanyl), —CH₂S(pyridinyl), —CH₂S(methylpyridinyl), —CH₂SCH₂CH₂(phenyl), —CH₂SCH₂CH₂(pyrazinyl),—CH₂SCH₂CH₂(pyridinyl), —CH₂S(O)(CH₂)₃CH₃, —CH₂S(O)₂(CH₂)₃CH₃,—CH₂S(O)(phenyl), —CH₂S(O)₂(phenyl), —(CH₂)₄OCH(CH₃)₂,—(CH₂)₄CH(CH₃)OCH₃, —(CH₂)₄C(CH₃)₂OCH₃, —(CH₂)₅N(CH₃)₂, —O(CH₂)₄₋₇CH₃,—OCH₂CH₂O(CH₂)₂₋₄CH₃, —OCH₂CH₂OCH₂CH(CH₃)₂, —OCH₂CH═CH(CH₂)₂₋₃CH₃,—OCH₂CH₂OCH₂CH═CH₂, —OCH₂CH₂OCH₂CH═CH(CH₃), —OCH₂CH₂OCH₂CH═C(CH₃)₂,—OCH₂CH₂OCH₂CH₂C≡CH, —OCH₂CH₂O(CH₂)₂₋₃CH(CH₃)₂, —OCH₂CH₂S(CH₂)₂CH₃,—OCH₂(cyclohexyl), —OCH₂(tetrahydropyranyl), —OCH₂(phenyl) wherein saidphenyl is substituted with zero to 1 substituent selectedfrom —CH₃, —CH₂CH₃, —OCH₃, —OCF₃,and —OCH₂CH₃; —OCH₂CH₂O(cyclohexyl), —OCH₂CH₂O(methyl phenyl),—OCH₂CH₂OCH₂(cyclobutyl), —OCH₂CH₂OCH₂(phenyl), —OCH₂CH₂OCH₂(thiazolyl),—OCH₂CH₂OCH₂(thiophenyl), —OC(O)(CH₂)₄CH₃, —OC(O)C(CH₃)₂(CH₂)₃CH₃,—OC(O)(phenyl), —OC(O)NH(CH₂)₃CH₃, —OC(O)NH(CH₂)₅CH₃,—OC(O)N(CH₃)(CH₂)₃CH₃, —OC(O)N(CH₃)(CH₂)₄CH₃, —NHC(O)NH(CH₂)₃CH₃,—C(CH₃)═N—O(CH₂)₃CH₃, —C(CH₃)═N—OCH₂(phenyl),—C(CH₃)═N—OCH₂(fluorophenyl), —C(CH₃)═N—OCH₂(methoxyphenyl),—C(CH₃)═N—OCH₂CH₂(phenyl), —OC(O)NH(CH₂)₃CH₃, —OC(O)NH(CH₂)₅CH₃,—OC(O)N(CH₃)(CH₂)₃₋₄CH₃, —NHC(O)NH(CH₂)₃CH₃, phenyl, or pyridinyl;R_(2b) is: (i) a 6-membered spiro-ring having one oxygen atom andsubstituted with zero or 1 substituent selected from —(CH₂)₃CH₃; or(ii)═N—O—(CH₂)₃CH₃, ═N—O—CH₂CH(CH₃)₂, ═N—OCH₂CH₂(phenyl), or═N—O—CH₂CH₂CH₂(phenyl); R_(a) is H or —OH; each R_(b) is independently Hor —CH₃; and each Re is independently H, Cl, I, or —CH₃; with theproviso that if R₂ is —(CH₂)₆CH₃, then at least one of R_(b) and Re isnot H. Included in this embodiment are compounds of Formula (Ic). Alsoincluded in this embodiment are compounds in which R₁ is —OH.

One embodiment provides at least one compound of Formula (II) or a saltthereof, wherein X₁, X₂, X₃, and R_(2a) are defined in the first aspect.Included in this embodiment are compounds in which R_(2a) is—(CH₂)₅₋₆CH₃ or —CH₂O(CH₂)₃₋₄CH₃. Included in this embodiment arecompounds of Formula (IIc). Also included in this embodiment arecompounds in which R₁ is —OH.

One embodiment provides at least one compound of Formula (III), Formula(IV), or Formula (V) or a salt thereof, wherein R₁, R₂, R_(2a) and R_(b)are defined in the first aspect.

Included in this embodiment are compounds in which R₂ is R_(2a).Included in this embodiment are compounds in which R₂ is R_(2a). Alsoincluded in this embodiment are compounds in which R₂ is R_(2a); R_(2a)is —(CH₂)₃CH₃, —(CH₂)₅CH₃, —(CH₂)₃(phenyl), or —C(O)(CH₂)₄CH₃; and eachR_(b) is —CH₃. Additionally, included in this embodiment are compoundsof Formula (IIIc), (IVc), and (Vc). Also included in this embodiment arecompounds in which R₁ is —OH.

One embodiment provides at least one compound of Formula (I) or a saltthereof, wherein R₁ is —OH or —OP(O)(OH)₂; R₂ is R_(2a); R_(2a)

is —(CH₂)₃CH₃, —(CH₂)₅CH₃, —CH₂CH═CHCH₂CH₃, —CH₂CH₂CH═CHCH₂CH₃,—(CH₂)₃CH═CHCH₃, —(CH₂)₃CH═C(CH₃)₂, —(CH₂)₄CH═CH₂, —(CH₂)₄CH═CHCH₃,—CH═CH(CH₂)₃CH₃, —CH═CH(CH₂)₃OCH₃, —CH═CHCH₂CH₂CH(CH₃)₂,—CH═CHCH₂CH₂CH₂OCH₃, —CH₂CH═CHCH═CHCH₃, —CH═CHCH₂CH₂CH═CH₂,—CH═CH(phenyl) wherein said phenyl is substituted with —CH₃or —OCH₃; —CH═CH(tetrahydropyranyl), —(CH₂)₁₋₃(phenyl) wherein saidphenyl is substituted with zero to 2 substituents independently selectedfrom F, I, —CH₃, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, and —CH₂C(O)N(CH₃)₂;—(CH₂)₂(methyl imidazolyl), —(CH₂)₂(methyl pyrazolyl),—(CH₂)₁₋₂(pyridinyl) wherein said pyridinyl is substituted with zero to1 substituent selectedfrom —OCH₃; —(CH₂)₂(pyrimidinyl), —(CH₂)₂(quinolinyl),—(CH₂)₂₋₃(tetrahydropyranyl), —CH₂O(CH₂)₃₋₄CH₃, —CH₂OCH₂CH₂CH(CH₃)₂,—CH₂OCH₂CH₂C(CH₃)₃, —CH₂O(CH₂)₉CH₃, —CH₂OCH₂CH₂CH₂CF₃,—CH₂OCH₂CH═CHCH₂CH₃, —CH₂OCH₂CH═C(CH₃)₂, —CH₂OCH₂CH═CHCH₂CH₂CH₃,—CH₂OCH₂CH₂CH═CH₂, —CH₂OCH₂CH₂CH₂CH═CH₂, —CH₂OCH₂CH₂CH═C(CH₃)₂,—CH₂OCH₂CH₂CH(OH)CH₃, —CH₂OCH₂CH₂CH₂CH₂OH, —CH₂OCH₂CH₂CH₂C(CH₃)₂(OH),—CH₂OCH₂CH₂OCH₃, —CH₂OCH₂CH₂CH₂OCH₃, —CH₂OCH₂CH₂OCH₂CH₂CH₃,—CH₂O(phenyl) wherein said phenyl is substituted with zero to 3substituents independently selected from F, Cl, —CH₃, —CH(CH₃)₂,—C(CH₃)₃, —OCH₃, —OCF₃, —(CH₂)₁₋₆OCH₃, —C(O)N(CH₃)₂, —CH₂N(CH₃)₂,—C(O)N(CH₂CH₃)(CH₃), —C(O)N(CH₃)(CH₂CH₂CH₂CH₃), and—C(O)N(CH₃)(CH₂CH(CH₃)₂); —CH₂O(methoxy pyridinyl),—CH₂O(tetrahydropyranyl), —CH₂O(trifluoromethyl, methyl pyrazolyl),—CH₂OCH₂(phenyl) wherein said phenyl is substituted with zero to 1substituent selected from —CH₃ and —OCH₃; —CH₂OCH₂(methyl pyrazolyl),—CH₂OCH₂(tetrahydropyranyl), —CH₂OCH₂(thiophenyl),—CH₂OCH₂(trifluoromethyl thiophenyl), —CH₂OCH₂(ethyl thiophenyl),—CH₂OCH₂(dimethyl thiophenyl), —CH₂CH₂OCH₂CH₃, —CH₂CH₂OCH₂CH(CH₃)₂,—CH₂CH₂O(methoxyphenyl), —CH₂CH₂OCH₂(cyclopropyl), —CH₂CH₂SCH(CH₃)₂,—(CH₂)₃OCH₂CH₃, —(CH₂)₃OCH(CH₃)₂, —(CH₂)₃OCH₂CH₂CH═CH₂,—(CH₂)₃O(oxetanyl), —(CH₂)₃O(tetramethyl cyclohexyl), —(CH₂)₃OCH₂SCH₃,—CH₂S(CH₂)₂₋₄CH₃, —CH₂SCH(CH₃)₂, —CH₂SCH₂CH(CH₃)₂, —CH₂SCH₂C(CH₃)₃,—CH₂SCH₂CH₂CH(CH₃)₂, —CH₂SCH₂CH₂C(CH₃)₃, —CH₂SCH₂CH₂Si(CH₃)₃,—CH₂CH₂S(CH₂)₁₋₂CH₃, —CH₂CH₂SCH₂CH(CH₃)₂, —CH₂S(phenyl) wherein saidphenyl is substituted with zero to 2 substituents independently selectedfrom —CH₃, —CH(CH₃)₂,and —OCH₃; —CH₂S(adamantanyl), —CH₂S(pyridinyl), —CH₂S(methylpyridinyl), —CH₂SCH₂CH₂(phenyl), —CH₂SCH₂CH₂(pyrazinyl),—CH₂SCH₂CH₂(pyridinyl), —CH₂S(O)(CH₂)₃CH₃, —CH₂S(O)₂(CH₂)₃CH₃,—CH₂S(O)(phenyl), —CH₂S(O)₂(phenyl), —(CH₂)₄OCH(CH₃)₂,—(CH₂)₄CH(CH₃)OCH₃, —(CH₂)₄C(CH₃)₂OCH₃, —(CH₂)₅N(CH₃)₂, —O(CH₂)₄₋₇CH₃,—OCH₂CH₂O(CH₂)₂₋₄CH₃, —OCH₂CH₂OCH₂CH(CH₃)₂, —OCH₂CH═CH(CH₂)₂₋₃CH₃,—OCH₂CH₂OCH₂CH═CH₂, —OCH₂CH₂OCH₂CH═CH(CH₃), —OCH₂CH₂OCH₂CH═C(CH₃)₂,—OCH₂CH₂OCH₂CH₂C≡CH, —OCH₂CH₂O(CH₂)₂₋₃CH(CH₃)₂, —OCH₂CH₂S(CH₂)₂CH₃,—OCH₂(cyclohexyl), —OCH₂(tetrahydropyranyl), —OCH₂(phenyl) wherein saidphenyl is substituted with zero to 1 substituent selectedfrom —CH₃, —CH₂CH₃, —OCH₃, —OCF₃,and —OCH₂CH₃; —OCH₂CH₂O(cyclohexyl), —OCH₂CH₂O(methyl phenyl),—OCH₂CH₂OCH₂(cyclobutyl), —OCH₂CH₂OCH₂(phenyl), —OCH₂CH₂OCH₂(thiazolyl),—OCH₂CH₂OCH₂(thiophenyl), —OC(O)(CH₂)₄CH₃, —OC(O)C(CH₃)₂(CH₂)₃CH₃,—OC(O)(phenyl), —OC(O)NH(CH₂)₃CH₃, —OC(O)NH(CH₂)₅CH₃,—OC(O)N(CH₃)(CH₂)₃CH₃, —OC(O)N(CH₃)(CH₂)₄CH₃, —NHC(O)NH(CH₂)₃CH₃,—C(CH₃)═N—O(CH₂)₃CH₃, —C(CH₃)═N—OCH₂(phenyl),—C(CH₃)═N—OCH₂(fluorophenyl), —C(CH₃)═N—OCH₂(methoxyphenyl),—C(CH₃)═N—OCH₂CH₂(phenyl), —OC(O)NH(CH₂)₃CH₃, —OC(O)NH(CH₂)₅CH₃,—OC(O)N(CH₃)(CH₂)₃₋₄CH₃, —NHC(O)NH(CH₂)₃CH₃, phenyl, or pyridinyl; R_(a)is H or —OH; each R_(b) is independently H or —CH₃; and each R, isindependently H, Cl, I, or —CH₃; with the proviso that if R₂ is—(CH₂)₆CH₃, then at least one of R_(b) and R, is not H. Included in thisembodiment are compounds of Formula (Ic). Also included in thisembodiment are compounds in which R₁ is —OH.

One embodiment provides at least one compound of Formula (I) or a saltthereof, wherein R₁ is —OH or —OP(O)(OH)₂; R₂ is R_(2b); R_(2b) is: (i)a 6-membered spiro-ring having one oxygen atom and substituted with zeroor 1 substituent selected from —(CH₂)₃CH₃; or (ii) ═N—O—(CH₂)₃CH₃,═N—O—CH₂CH(CH₃)₂, ═N—OCH₂CH₂(phenyl), or ═N—O—CH₂CH₂CH₂(phenyl); R_(a)is H or —OH; each R_(b) is independently H or —CH₃; and each R, isindependently H, Cl, I, or —CH₃. Included in this embodiment arecompounds of Formula (Ic). Also included in this embodiment arecompounds in which R₁ is —OH.

One embodiment provides at least one compound of Formula (I) or a saltthereof, wherein R₂ is R_(2a); R_(2a)

is —(CH₂)₃₋₆CH₃, —(CH₂)₁₋₄CH═CR_(x)R_(x), —(CH₂)₁₋₄CH═CR_(x)(CH₂CH₃),—CH═CH(CH₂)₁₋₃C(R_(x))₃, —CH═CH(CH₂)₁₋₃OCH₃,—(CH₂)₁₋₃CH═CHCH═CR_(x)R_(x), —CH═CH(CH₂)₁₋₃CH═CR_(x)R_(x), —CH═CHR_(z),—(CH₂)₁₋₃R_(z), —(CH₂)₁₋₃O(CH₂)₀₋₃R_(z), —(CH₂)₁₋₃S(CH₂)₀₋₃R_(z),—CH₂S(O)R_(z), —CH₂S(O)₂R_(z), —O(CH₂)₁₋₂R_(z),—O(CH₂)₁₋₂O(CH₂)₀₋₂R_(z), —OC(O)R_(z), —(CH₂)₁₋₄O(CH₂)₀₋₉C(R_(x))₃,—(CH₂)₁₋₄O(CH₂)₀₋₉CF₃, —(CH₂)₁₋₄CR_(x)R_(x)O(CH₂)₀₋₄C(R_(x))₃,—(CH₂)₁₋₃O(CH₂)₁₋₄CH═CR_(x)(CH₂)₀₋₃CH₃,—(CH₂)₁₋₃O(CH₂)₁₋₄CH═CR_(x)R_(x), —(CH₂)₁₋₃O(CH₂)₁₋₄C(OH)R_(x)R_(x),—(CH₂)₁₋₃O(CH₂)₁₋₄O(CH₂)₀₋₃CH₃, —(CH₂)₁₋₃S(CH₂)₀₋₄C(R_(x))₃,—(CH₂)₀₋₃O(CH₂)₁₋₄S(CH₂)₀₋₃C(R_(x))₃, —(CH₂)₁₋₃S(CH₂)₁₋₄Si(CH₃)₃,—(CH₂)₁₋₃S(O)(CH₂)₀₋₄C(R_(x))₃, —(CH₂)₁₋₃S(O)₂(CH₂)₀₋₄C(R_(x))₃,—(CH₂)₁₋₅NR_(x)R_(x), or —O(CH₂)₁₋₇C(R_(x))₃; and R₁, R_(a), R_(b), andR, are defined in the first aspect. Included in this embodiment arecompounds in which R_(2a)is —(CH₂)₃CH₃, —(CH₂)₅₋₆CH₃, —CH₂CH═CHCH₂CH₃, —CH₂CH₂CH═CHCH₂CH₃,—(CH₂)₃CH═CHCH₃, —(CH₂)₃CH═C(CH₃)₂, —(CH₂)₄CH═CH₂, —(CH₂)₄CH═CHCH₃,—CH═CH(CH₂)₃CH₃, —CH═CH(CH₂)₃OCH₃, —CH═CHCH₂CH₂CH(CH₃)₂,—CH═CHCH₂CH₂CH₂OCH₃, —CH₂CH═CHCH═CHCH₃, —CH═CHCH₂CH₂CH═CH₂,—CH═CH(phenyl) wherein said phenyl is substituted with —CH₃or —OCH₃; —CH═CH(tetrahydropyranyl), —(CH₂)₁₋₃(phenyl) wherein saidphenyl is substituted with zero to 2 substituents independently selectedfrom F, I, —CH₃, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, and —CH₂C(O)N(CH₃)₂;—(CH₂)₂(methyl imidazolyl), —(CH₂)₂(methyl pyrazolyl),—(CH₂)₁₋₂(pyridinyl) wherein said pyridinyl is substituted with zero to1 substituent selectedfrom —OCH₃; —(CH₂)₂(pyrimidinyl), —(CH₂)₂(quinolinyl),—(CH₂)₂₋₃(tetrahydropyranyl), —CH₂O(CH₂)₃₋₄CH₃, —CH₂OCH₂CH₂CH(CH₃)₂,—CH₂OCH₂CH₂C(CH₃)₃, —CH₂O(CH₂)₉CH₃, —CH₂OCH₂CH₂CH₂CF₃,—CH₂OCH₂CH═CHCH₂CH₃, —CH₂OCH₂CH═C(CH₃)₂, —CH₂OCH₂CH═CHCH₂CH₂CH₃,—CH₂OCH₂CH₂CH═CH₂, —CH₂OCH₂CH₂CH₂CH═CH₂, —CH₂OCH₂CH₂CH═C(CH₃)₂,—CH₂OCH₂CH₂CH(OH)CH₃, —CH₂OCH₂CH₂CH₂CH₂OH, —CH₂OCH₂CH₂CH₂C(CH₃)₂(OH),—CH₂OCH₂CH₂OCH₃, —CH₂OCH₂CH₂CH₂OCH₃, —CH₂OCH₂CH₂OCH₂CH₂CH₃,—CH₂O(phenyl) wherein said phenyl is substituted with zero to 3substituents independently selected from F, Cl, —CH₃, —CH(CH₃)₂,—C(CH₃)₃, —OCH₃, —OCF₃, —(CH₂)₁₋₆OCH₃, —C(O)N(CH₃)₂, —CH₂N(CH₃)₂,—C(O)N(CH₂CH₃)(CH₃), —C(O)N(CH₃)(CH₂CH₂CH₂CH₃), and—C(O)N(CH₃)(CH₂CH(CH₃)₂); —CH₂O(methoxy pyridinyl),—CH₂O(tetrahydropyranyl), —CH₂O(trifluoromethyl, methyl pyrazolyl),—CH₂OCH₂(phenyl) wherein said phenyl is substituted with zero to 1substituent selected from —CH₃ and —OCH₃; —CH₂OCH₂(methyl pyrazolyl),—CH₂OCH₂(tetrahydropyranyl), —CH₂OCH₂(thiophenyl), —CH₂OCH₂(trifluoromethyl thiophenyl), —CH₂OCH₂(ethyl thiophenyl), —CH₂OCH₂(dimethylthiophenyl), —CH₂CH₂OCH₂CH₃, —CH₂CH₂OCH₂CH(CH₃)₂,—CH₂CH₂O(methoxyphenyl), —CH₂CH₂OCH₂(cyclopropyl), —CH₂CH₂SCH(CH₃)₂,—(CH₂)₃OCH₂CH₃, —(CH₂)₃OCH(CH₃)₂, —(CH₂)₃OCH₂CH₂CH═CH₂,—(CH₂)₃O(oxetanyl), —(CH₂)₃O(tetramethyl cyclohexyl), —(CH₂)₃OCH₂SCH₃,—CH₂S(CH₂)₂₋₄CH₃, —CH₂SCH(CH₃)₂, —CH₂SCH₂CH(CH₃)₂, —CH₂SCH₂C(CH₃)₃,—CH₂SCH₂CH₂CH(CH₃)₂, —CH₂SCH₂CH₂C(CH₃)₃, —CH₂SCH₂CH₂Si(CH₃)₃,—CH₂CH₂S(CH₂)₁₋₂CH₃, —CH₂CH₂SCH₂CH(CH₃)₂, —CH₂S(phenyl) wherein saidphenyl is substituted with zero to 2 substituents independently selectedfrom —CH₃, —CH(CH₃)₂,and —OCH₃; —CH₂S(adamantanyl), —CH₂S(pyridinyl), —CH₂S(methylpyridinyl), —CH₂SCH₂CH₂(phenyl), —CH₂SCH₂CH₂(pyrazinyl),—CH₂SCH₂CH₂(pyridinyl), —CH₂S(O)(CH₂)₃CH₃, —CH₂S(O)₂(CH₂)₃CH₃,—CH₂S(O)(phenyl), —CH₂S(O)₂(phenyl), —(CH₂)₄OCH(CH₃)₂,—(CH₂)₄CH(CH₃)OCH₃, —(CH₂)₄C(CH₃)₂OCH₃, or —(CH₂)₅N(CH₃)₂.

One embodiment provides at least one compound of Formula (I) or a saltthereof, wherein —O(CH₂)₁₋₄O(CH₂)₀₋₄C(R_(x))₃,—O(CH₂)₁₋₄CH═CR_(x)(CH₂)₀₋₃CH₃, —O(CH₂)₁₋₄O(CH₂)₀₋₃C(R_(x))₃,—O(CH₂)₁₋₄O(CH₂)₁₋₃CH═CR_(x)R_(x), or —O(CH₂)₁₋₄O(CH₂)₁₋₃C≡CR_(x); andR₁, R_(a), R_(b), and R, are defined in the first aspect. Included inthis embodiment are compounds in which R_(2a)

is —O(CH₂)₄-7CH₃, —OCH₂CH₂O(CH₂)₂₋₄CH₃, —OCH₂CH₂OCH₂CH(CH₃)₂,—OCH₂CH═CH(CH₂)₂₋₃CH₃, —OCH₂CH₂OCH₂CH═CH₂, —OCH₂CH₂OCH₂CH═CH(CH₃),—OCH₂CH₂OCH₂CH═C(CH₃)₂, —OCH₂CH₂OCH₂CH₂C≡CH, —OCH₂CH₂O(CH₂)₂₋₃CH(CH₃)₂,—OCH₂CH₂S(CH₂)₂CH₃, —OCH₂(cyclohexyl), —OCH₂(tetrahydropyranyl),—OCH₂(phenyl) wherein said phenyl is substituted with zero to 1substituent selectedfrom —CH₃, —CH₂CH₃, —OCH₃, —OCF₃,and —OCH₂CH₃; —OCH₂CH₂O(cyclohexyl), —OCH₂CH₂O(methyl phenyl),—OCH₂CH₂OCH₂(cyclobutyl), —OCH₂CH₂OCH₂(phenyl), —OCH₂CH₂OCH₂(thiazolyl),or —OCH₂CH₂OCH₂(thiophenyl).

One embodiment provides at least one compound of Formula (I) or a saltthereof, —C(O)(CH₂)₀₋₄C(R_(x))₃, —OC(O)(CH₂)₀₋₄C(R_(x))₃,—OC(O)CR_(x)R_(x)(CH₂)₀₋₄C(R_(x))₃, —OC(O)NR_(x)(CH₂)₀₋₅C(R_(x))₃,—NR_(x)C(O)NR_(x)(CH₂)₀₋₅C(R_(x))₃, —C(CH₃)═N—O(CH₂)₀₋₅C(R_(x))₃,—C(CH₃)═N—O(CH₂)₁₋₂(phenyl), —C(CH₃)═N—O(CH₂)₁₋₂(fluorophenyl),—C(CH₃)═N—O(CH₂)₁₋₂(methoxyphenyl), phenyl, or pyridinyl; and R₁, R_(a),R_(b), and R, are defined in the first aspect. Included in thisembodiment are compounds in which R_(2a)

is —C(O)(CH₂)₄CH₃, —OC(O)(CH₂)₄CH₃, —OC(O)C(CH₃)₂(CH₂)₃CH₃,—OC(O)(phenyl), —O C(O)NH(CH₂)₃CH₃, —OC(O)NH(CH₂)₅CH₃,—OC(O)N(CH₃)(CH₂)₃CH₃, —OC(O)N(CH₃)(CH₂)₄CH₃, —NHC(O)NH(CH₂)₃CH₃,—C(CH₃)═N—O(CH₂)₃CH₃, —C(CH₃)═N—OCH₂(phenyl),—C(CH₃)═N—OCH₂(fluorophenyl), —C(CH₃)═N—OCH₂(methoxyphenyl),—C(CH₃)═N—OCH₂CH₂(phenyl), —OC(O)NH(CH₂)₃CH₃, —OC(O)NH(CH₂)₅CH₃,—OC(O)N(CH₃)(CH₂)₃₋₄CH₃, —NHC(O)NH(CH₂)₃CH₃, phenyl, or pyridinyl.

One embodiment provides at least one compound of Formula (I) or a saltthereof, wherein R₁ is —OH or —OP(O)(OH)₂; R₂ is R_(2b); R_(2b) is a6-membered spiro-ring having one oxygen atom and substituted with zeroor 1 substituent selected from —(CH₂)₃CH₃; R_(a) is H or —OH; each R_(b)is independently H or —CH₃; and each Re is independently H, Cl, I, or—CH₃. Included in this embodiment are compounds of Formula (Ic). Alsoincluded in this embodiment are compounds in which R₁ is —OH.Additionally, included in this embodiment are compounds having thefollowing structures:

wherein Ry is H or —(CH₂)₃CH₃.

One embodiment provides at least one compound of Formula (I) or a saltthereof, wherein R₁ is —OH or —OP(O)(OH)₂; R₂ is R_(2b); R_(2b) is═N—O—(CH₂)₃CH₃, ═N—O—CH₂CH(CH₃)₂, ═N—OCH₂CH₂(phenyl), or═N—O—CH₂CH₂CH₂(phenyl); R_(a) is H or —OH; each R_(b) is independently Hor —CH₃; and each Re is independently H, Cl, I, or —CH₃. Included inthis embodiment are compounds of Formula (Ic). Also included in thisembodiment are compounds in which R₁ is —OH.

One embodiment provides at least one compound of Formula (I) or a saltthereof having the structure:

wherein: R₁ is —OH or —OP(O)(OH)₂; and R₂is —(CH₂)₅₀CH₃, —(CH₂)₃OCH₂CH₃, —CH₂O(methoxyphenyl),or —CH₂CH₂(methoxyphenyl).

One embodiment provides at least one compound of Formula (I) or a saltthereof having the structure:

wherein R₁ is —OH or —OP(O)(OH)₂; and R₂ is —(CH₂)₅OCH₃ or—(CH₂)₃OCH₂CH₃. Included in this embodiment are compounds having thestructures:

One embodiment provides at least one compound of Formula (I) or a saltthereof having the structure:

wherein R₁ is —OH or —OP(O)(OH)₂; and R₂ is —CH₂O(methoxyphenyl) or—CH₂CH₂(methoxyphenyl). Included in this embodiment are compounds havingthe structures:

One embodiment provides a compound selected from((1R,3S)-1-amino-3-((S)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(672);((1R,3S)-1-amino-3-((R)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol (673);((1R,3R)-1-amino-3-(6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(674);((1R,3R)-1-amino-3-((S)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol;((1R,3R)-1-amino-3-((R)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol;((1R,3S)-1-amino-3-((R)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(678);((1R,3S)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(679);((1R,3R)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol;and salts thereof.

One embodiment provides a compound selected from((1R,3S)-1-amino-3-((R)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3S)-1-amino-3-((S)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3R)-1-amino-3-((R)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3R)-1-amino-3-((S)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3S)-1-amino-3-((R)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3S)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate; and salts thereof.

One embodiment provides a compound selected from((1R,3S)-1-amino-3-((S)-6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol (676);((1R,3S)-1-amino-3-((R)-6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(677);((1R,3S)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(681);((1R,3S)-1-amino-3-((R)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(682);((1R,3R)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol (683);((1R,3S)-1-amino-3-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(684);((1R,3S)-1-amino-3-((S)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(685);((1R,3R)-1-amino-3-(6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol;((1R,3R)-1-amino-3-((S)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol;((1R,3R)-1-amino-3-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol;and salts thereof.

One embodiment provides a compound selected from((1R,3R)-1-amino-3-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate (688);((1R,3S)-1-amino-3-((R)-6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3S)-1-amino-3-((S)-6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3S)-1-amino-3-((R)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3S)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3R)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate (698);((1R,3S)-1-amino-3-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3S)-1-amino-3-((S)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3R)-1-amino-3-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate;((1R,3R)-1-amino-3-((S)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate; and salts thereof.

The present invention may be embodied in other specific forms withoutdeparting from the spirit or essential attributes thereof. Thisinvention encompasses all combinations of the aspects and/or embodimentsof the invention noted herein. It is understood that any and allembodiments of the present invention may be taken in conjunction withany other embodiment or embodiments to describe additional embodiments.It is also to be understood that each individual element of theembodiments is meant to be combined with any and all other elements fromany embodiment to describe an additional embodiment.

Definitions

The features and advantages of the invention may be more readilyunderstood by those of ordinary skill in the art upon reading thefollowing detailed description. It is to be appreciated that certainfeatures of the invention that are, for clarity reasons, described aboveand below in the context of separate embodiments, may also be combinedto form a single embodiment. Conversely, various features of theinvention that are, for brevity reasons, described in the context of asingle embodiment, may also be combined so as to form sub-combinationsthereof. Embodiments identified herein as exemplary or preferred areintended to be illustrative and not limiting.

Unless specifically stated otherwise herein, references made in thesingular may also include the plural. For example, “a” and “an” mayrefer to either one, or one or more.

As used herein, the phase “compounds and/or salts thereof” refers to atleast one compound, at least one salt of the compounds, or a combinationthereof. For example, compounds of Formula (I) and/or salts thereofincludes a compound of Formula (I); two compounds of Formula (I); a saltof a compound of Formula (I); a compound of Formula (I) and one or moresalts of the compound of Formula (I); and two or more salts of acompound of Formula (I).

Unless otherwise indicated, any atom with unsatisfied valences isassumed to have hydrogen atoms sufficient to satisfy the valences.

The definitions set forth herein take precedence over definitions setforth in any patent, patent application, and/or patent applicationpublication incorporated herein by reference.

Listed below are definitions of various terms used to describe thepresent invention. These definitions apply to the terms as they are usedthroughout the specification (unless they are otherwise limited inspecific instances) either individually or as part of a larger group.

Throughout the specification, groups and substituents thereof may bechosen by one skilled in the field to provide stable moieties andcompounds.

In accordance with a convention used in the art,

is used in structural formulas herein to depict the bond that is thepoint of attachment of the moiety or substituent to the core or backbonestructure.

The term “alkyl” as used herein, refers to both branched andstraight-chain saturated aliphatic hydrocarbon groups containing, forexample, from 1 to 12 carbon atoms, from 1 to 6 carbon atoms, and from 1to 4 carbon atoms. Examples of alkyl groups include, but are not limitedto, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and i-propyl), butyl(e.g., n-butyl, i-butyl, sec-butyl, and t-butyl), and pentyl (e.g.,n-pentyl, isopentyl, neopentyl), n-hexyl, 2-methylpentyl, 2-ethylbutyl,3-methylpentyl, and 4-methylpentyl. When numbers appear in a subscriptafter the symbol “C”, the subscript defines with more specificity thenumber of carbon atoms that a particular group may contain. For example,“C₁₋₄ alkyl” denotes straight and branched chain alkyl groups with oneto four carbon atoms.

As used herein, “alkylene” refers to a bivalent alkyl radical having thegeneral formula —(CH₂)_(n)—, where n is the number of methylene units.Non-limiting examples include methylene, dimethylene, trimethylene,tetramethylene, pentamethylene, and hexamethylene. For example,“(CH₂)₁₋₆” denotes straight chain alkylene groups with one to six carbonatoms. Further, for example, “(CH₂)₀₋₄” denotes a bond and straightchain alkylene groups with one to four carbon atoms.

The term “cycloalkyl,” as used herein, refers to a group derived from anon-aromatic monocyclic or polycyclic hydrocarbon molecule by removal ofone hydrogen atom from a saturated ring carbon atom. Representativeexamples of cycloalkyl groups include, but are not limited to,cyclopropyl, cyclopentyl, and cyclohexyl. When numbers appear in asubscript after the symbol “C”, the subscript defines with morespecificity the number of carbon atoms that a particular cycloalkylgroup may contain. For example, “C₃₋₆ cycloalkyl” denotes cycloalkylgroups with three to six carbon atoms.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The compounds of Formulas (I), (II), (III), (IV), and (V) can form saltswhich are also within the scope of this invention. Unless otherwiseindicated, reference to an inventive compound is understood to includereference to one or more salts thereof. The term “salt(s)” denotesacidic and/or basic salts formed with inorganic and/or organic acids andbases. In addition, the term “salt(s) may include zwitterions (innersalts), e.g., when a compound of Formulas (I), (II), (III), (IV), or (V)contains both a basic moiety, such as an amine or a pyridine orimidazole ring, and an acidic moiety, such as a carboxylic acid.Pharmaceutically acceptable (i.e., non-toxic, physiologicallyacceptable) salts are preferred, such as, for example, acceptable metaland amine salts in which the cation does not contribute significantly tothe toxicity or biological activity of the salt. However, other saltsmay be useful, e.g., in isolation or purification steps which may beemployed during preparation, and thus, are contemplated within the scopeof the invention. Salts of the compounds of the Formulas (I), (II),(III), (IV), or (V) may be formed, for example, by reacting a compoundof the Formulas (I), (II), (III), (IV), or (V) with an amount of acid orbase, such as an equivalent amount, in a medium such as one in which thesalt precipitates or in an aqueous medium followed by lyophilization.

Exemplary acid addition salts include acetates (such as those formedwith acetic acid or trihaloacetic acid, for example, trifluoroaceticacid), adipates, alginates, ascorbates, aspartates, benzoates,benzenesulfonates, bisulfates, borates, butyrates, citrates,camphorates, camphorsulfonates, cyclopentanepropionates, digluconates,dodecylsulfates, ethanesulfonates, fumarates, glucoheptanoates,glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides(formed with hydrochloric acid), hydrobromides (formed with hydrogenbromide), hydroiodides, maleates (formed with maleic acid),2-hydroxyethanesulfonates, lactates, methanesulfonates (formed withmethanesulfonic acid), 2-naphthalenesulfonates, nicotinates, nitrates,oxalates, pectinates, persulfates, 3-phenylpropionates, phosphates,picrates, pivalates, propionates, salicylates, succinates, sulfates(such as those formed with sulfuric acid), sulfonates (such as thosementioned herein), tartrates, thiocyanates, toluenesulfonates such astosylates, undecanoates, and the like.

Exemplary basic salts include ammonium salts, alkali metal salts such assodium, lithium, and potassium salts; alkaline earth metal salts such ascalcium and magnesium salts; barium, zinc, and aluminum salts; saltswith organic bases (for example, organic amines) such as trialkylaminessuch as triethylamine, procaine, dibenzylamine,N-benzyl-Q-phenethylamine, 1-ephenamine, N,N′-dibenzylethylene-diamine,dehydroabietylamine, N-ethylpiperidine, benzylamine, dicyclohexylamineor similar pharmaceutically acceptable amines and salts with amino acidssuch as arginine, lysine and the like. Basic nitrogen-containing groupsmay be quaternized with agents such as lower alkyl halides (e.g.,methyl, ethyl, propyl, and butyl chlorides, bromides and iodides),dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl, and diamylsulfates), long chain halides (e.g., decyl, lauryl, myristyl and stearylchlorides, bromides and iodides), aralkyl halides (e.g., benzyl andphenethyl bromides), and others. Preferred salts includemonohydrochloride, hydrogensulfate, methanesulfonate, phosphate ornitrate salts.

The compounds of Formulas (I), (II), (III), (IV), and (V) can beprovided as amorphous solids or crystalline solids. Lyophilization canbe employed to provide the compounds as a solid.

It should further be understood that solvates (e.g., hydrates) of thecompounds of Formulas (I), (II), (III), (IV), and (V) are also withinthe scope of the present invention. The term “solvate” means a physicalassociation of a compound of Formulas (I), (II), (III), (IV), or (V)with one or more solvent molecules, whether organic or inorganic. Thisphysical association includes hydrogen bonding. In certain instances thesolvate will be capable of isolation, for example when one or moresolvent molecules are incorporated in the crystal lattice of thecrystalline solid. “Solvate” encompasses both solution-phase andisolable solvates. Exemplary solvates include hydrates, ethanolates,methanolates, isopropanolates, acetonitrile solvates, and ethyl acetatesolvates. Methods of solvation are known in the art.

In addition, compounds of Formulas (I), (II), (III), (IV), and (V),subsequent to their preparation, can be isolated and purified to obtaina composition containing an amount by weight equal to or greater than99% of a compound of Formulas (I), (II), (III), (IV), and (V)(“substantially pure”), which is then used or formulated as describedherein.

Such “substantially pure” compounds of Formulas (I), (II), (III), (IV),and (V) are also contemplated herein as part of the present invention.

“Stable compound” and “stable structure” are meant to indicate acompound that is sufficiently robust to survive isolation to a usefuldegree of purity from a reaction mixture, and formulation into anefficacious therapeutic agent. The present invention is intended toembody stable compounds.

“Therapeutically effective amount” is intended to include an amount of acompound of the present invention alone or an amount of the combinationof compounds claimed or an amount of a compound of the present inventionin combination with other active ingredients effective to act as anagonist to S1P₁, or effective to treat or prevent autoimmune and/orinflammatory disease states, such as multiple sclerosis and rheumatoidarthritis.

As used herein, “treating” or “treatment” cover the treatment of adisease-state in a mammal, particularly in a human, and include: (a)preventing the disease-state from occurring in a mammal, in particular,when such mammal is predisposed to the disease-state but has not yetbeen diagnosed as having it; (b) inhibiting the disease-state, i.e.,arresting its development; and/or (c) relieving the disease-state, i.e.,causing regression of the disease state.

The compounds of the present invention are intended to include allisotopes of atoms occurring in the present compounds. Isotopes includethose atoms having the same atomic number but different mass numbers. Byway of general example and without limitation, isotopes of hydrogeninclude deuterium (D) and tritium (T). Isotopes of carbon include ¹³Cand ¹⁴C. Isotopically-labeled compounds of the invention can generallybe prepared by conventional techniques known to those skilled in the artor by processes analogous to those described herein, using anappropriate isotopically-labeled reagent in place of the non-labeledreagent otherwise employed. For example, methyl (—CH₃) also includesdeuterated methyl groups such as —CD₃.

Compounds in accordance with Formulas (I), (II), (III), (IV), and (V)and/or pharmaceutically acceptable salts thereof can be administered byany means suitable for the condition to be treated, which can depend onthe need for site-specific treatment or quantity of Formula (I) compoundto be delivered.

Also embraced within this invention is a class of pharmaceuticalcompositions comprising a compound of Formulas (I), (II), (III), (IV),or (V) and/or pharmaceutically acceptable salts thereof; and one or morenon-toxic, pharmaceutically-acceptable carriers and/or diluents and/oradjuvants (collectively referred to herein as “carrier” materials) and,if desired, other active ingredients. The compounds of Formulas (I),(II), (III), (IV), and (V) may be administered by any suitable route,preferably in the form of a pharmaceutical composition adapted to such aroute, and in a dose effective for the treatment intended. The compoundsand compositions of the present invention may, for example, beadministered orally, mucosally, or parentally including intravascularly,intravenously, intraperitoneally, subcutaneously, intramuscularly, andintrasternally in dosage unit formulations containing conventionalpharmaceutically acceptable carriers, adjuvants, and vehicles. Forexample, the pharmaceutical carrier may contain a mixture of mannitol orlactose and microcrystalline cellulose. The mixture may containadditional components such as a lubricating agent, e.g. magnesiumstearate and a disintegrating agent such as crospovidone. The carriermixture may be filled into a gelatin capsule or compressed as a tablet.The pharmaceutical composition may be administered as an oral dosageform or an infusion, for example.

For oral administration, the pharmaceutical composition may be in theform of, for example, a tablet, capsule, liquid capsule, suspension, orliquid. The pharmaceutical composition is preferably made in the form ofa dosage unit containing a particular amount of the active ingredient.For example, the pharmaceutical composition may be provided as a tabletor capsule comprising an amount of active ingredient in the range offrom about 0.1 to 1000 mg, preferably from about 0.25 to 250 mg, andmore preferably from about 0.5 to 100 mg. A suitable daily dose for ahuman or other mammal may vary widely depending on the condition of thepatient and other factors, but, can be determined using routine methods.

Any pharmaceutical composition contemplated herein can, for example, bedelivered orally via any acceptable and suitable oral preparations.Exemplary oral preparations, include, but are not limited to, forexample, tablets, troches, lozenges, aqueous and oily suspensions,dispersible powders or granules, emulsions, hard and soft capsules,liquid capsules, syrups, and elixirs. Pharmaceutical compositionsintended for oral administration can be prepared according to anymethods known in the art for manufacturing pharmaceutical compositionsintended for oral administration. In order to provide pharmaceuticallypalatable preparations, a pharmaceutical composition in accordance withthe invention can contain at least one agent selected from sweeteningagents, flavoring agents, coloring agents, demulcents, antioxidants, andpreserving agents.

A tablet can, for example, be prepared by admixing at least one compoundof Formulas (I), (II), (III), (IV), or (V) and/or at least onepharmaceutically acceptable salt thereof with at least one non-toxicpharmaceutically acceptable excipient suitable for the manufacture oftablets. Exemplary excipients include, but are not limited to, forexample, inert diluents, such as, for example, calcium carbonate, sodiumcarbonate, lactose, calcium phosphate, and sodium phosphate; granulatingand disintegrating agents, such as, for example, microcrystallinecellulose, sodium crosscarmellose, corn starch, and alginic acid;binding agents, such as, for example, starch, gelatin,polyvinyl-pyrrolidone, and acacia; and lubricating agents, such as, forexample, magnesium stearate, stearic acid, and talc. Additionally, atablet can either be uncoated, or coated by known techniques to eithermask the bad taste of an unpleasant tasting drug, or delaydisintegration and absorption of the active ingredient in thegastrointestinal tract thereby sustaining the effects of the activeingredient for a longer period. Exemplary water soluble taste maskingmaterials, include, but are not limited to,hydroxypropyl-methylcellulose and hydroxypropyl-cellulose. Exemplarytime delay materials, include, but are not limited to, ethyl celluloseand cellulose acetate butyrate.

Hard gelatin capsules can, for example, be prepared by mixing at leastone compound of Formulas (I), (II), (III), (IV), or (V) and/or at leastone salt thereof with at least one inert solid diluent, such as, forexample, calcium carbonate; calcium phosphate; and kaolin.

Soft gelatin capsules can, for example, be prepared by mixing at leastone compound of Formulas (I), (II), (III), (IV), or (V) and/or at leastone pharmaceutically acceptable salt thereof with at least one watersoluble carrier, such as, for example, polyethylene glycol; and at leastone oil medium, such as, for example, peanut oil, liquid paraffin, andolive oil.

An aqueous suspension can be prepared, for example, by admixing at leastone compound of Formulas (I), (II), (III), (IV), or (V) and/or at leastone pharmaceutically acceptable salt thereof with at least one excipientsuitable for the manufacture of an aqueous suspension. Exemplaryexcipients suitable for the manufacture of an aqueous suspension,include, but are not limited to, for example, suspending agents, suchas, for example, sodium carboxymethylcellulose, methylcellulose,hydroxypropylmethyl-cellulose, sodium alginate, alginic acid,polyvinyl-pyrrolidone, gum tragacanth, and gum acacia; dispersing orwetting agents, such as, for example, a naturally-occurring phosphatide,e.g., lecithin; condensation products of alkylene oxide with fattyacids, such as, for example, polyoxyethylene stearate; condensationproducts of ethylene oxide with long chain aliphatic alcohols, such as,for example heptadecaethylene-oxycetanol; condensation products ofethylene oxide with partial esters derived from fatty acids and hexitol,such as, for example, polyoxyethylene sorbitol monooleate; andcondensation products of ethylene oxide with partial esters derived fromfatty acids and hexitol anhydrides, such as, for example, polyethylenesorbitan monooleate. An aqueous suspension can also contain at least onepreservative, such as, for example, ethyl and n-propylp-hydroxybenzoate; at least one coloring agent; at least one flavoringagent; and/or at least one sweetening agent, including but not limitedto, for example, sucrose, saccharin, and aspartame.

Oily suspensions can, for example, be prepared by suspending at leastone compound of Formulas (I), (II), (III), (IV), or (V) and/or at leastone pharmaceutically acceptable salt thereof in either a vegetable oil,such as, for example, arachis oil; olive oil; sesame oil; and coconutoil; or in mineral oil, such as, for example, liquid paraffin. An oilysuspension can also contain at least one thickening agent, such as, forexample, beeswax; hard paraffin; and cetyl alcohol. In order to providea palatable oily suspension, at least one of the sweetening agentsalready described hereinabove, and/or at least one flavoring agent canbe added to the oily suspension. An oily suspension can further containat least one preservative, including, but not limited to, for example,an anti-oxidant, such as, for example, butylated hydroxyanisol, andalpha-tocopherol.

Dispersible powders and granules can, for example, be prepared byadmixing at least one compound of Formulas (I), (II), (III), (IV), or(V) and/or at least one pharmaceutically acceptable salt thereof with atleast one dispersing and/or wetting agent; at least one suspendingagent; and/or at least one preservative. Suitable dispersing agents,wetting agents, and suspending agents are as already described above.Exemplary preservatives include, but are not limited to, for example,anti-oxidants, e.g., ascorbic acid. In addition, dispersible powders andgranules can also contain at least one excipient, including, but notlimited to, for example, sweetening agents; flavoring agents; andcoloring agents.

An emulsion of at least one compound of Formulas (I), (II), (III), (IV),or (V) and/or at least one pharmaceutically acceptable salt thereof can,for example, be prepared as an oil-in-water emulsion. The oily phase ofthe emulsions comprising compounds of Formulas (I), (II), (III), (IV),or (V) may be constituted from known ingredients in a known manner. Theoil phase can be provided by, but is not limited to, for example, avegetable oil, such as, for example, olive oil and arachis oil; amineral oil, such as, for example, liquid paraffin; and mixturesthereof. While the phase may comprise merely an emulsifier, it maycomprise a mixture of at least one emulsifier with a fat or an oil orwith both a fat and an oil. Suitable emulsifying agents include, but arenot limited to, for example, naturally-occurring phosphatides, e.g., soybean lecithin; esters or partial esters derived from fatty acids andhexitol anhydrides, such as, for example, sorbitan monooleate; andcondensation products of partial esters with ethylene oxide, such as,for example, polyoxyethylene sorbitan monooleate. Preferably, ahydrophilic emulsifier is included together with a lipophilic emulsifierwhich acts as a stabilizer. It is also preferred to include both an oiland a fat. Together, the emulsifier(s) with or without stabilizer(s)make-up the so-called emulsifying wax, and the wax together with the oiland fat make up the so-called emulsifying ointment base which forms theoily dispersed phase of the cream formulations. An emulsion can alsocontain a sweetening agent, a flavoring agent, a preservative, and/or anantioxidant. Emulsifiers and emulsion stabilizers suitable for use inthe formulation of the present invention include Tween 60, Span 80,cetostearyl alcohol, myristyl alcohol, glyceryl monostearate, sodiumlauryl sulfate, glyceryl distearate alone or with a wax, or othermaterials well known in the art.

The compounds of Formulas (I), (II), (III), (IV), or (V) and/or at leastone pharmaceutically acceptable salt thereof can, for example, also bedelivered intravenously, subcutaneously, and/or intramuscularly via anypharmaceutically acceptable and suitable injectable form. Exemplaryinjectable forms include, but are not limited to, for example, sterileaqueous solutions comprising acceptable vehicles and solvents, such as,for example, water, Ringer's solution, and isotonic sodium chloridesolution; sterile oil-in-water microemulsions; and aqueous or oleaginoussuspensions.

Formulations for parenteral administration may be in the form of aqueousor non-aqueous isotonic sterile injection solutions or suspensions.These solutions and suspensions may be prepared from sterile powders orgranules using one or more of the carriers or diluents mentioned for usein the formulations for oral administration or by using other suitabledispersing or wetting agents and suspending agents. The compounds may bedissolved in water, polyethylene glycol, propylene glycol, ethanol, cornoil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodiumchloride, tragacanth gum, and/or various buffers. Other adjuvants andmodes of administration are well and widely known in the pharmaceuticalart. The active ingredient may also be administered by injection as acomposition with suitable carriers including saline, dextrose, or water,or with cyclodextrin (i.e. Captisol), cosolvent solubilization (i.e.propylene glycol) or micellar solubilization (i.e. Tween 80).

The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally acceptable diluent orsolvent, for example as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution, and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employed,including synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

A sterile injectable oil-in-water microemulsion can, for example, beprepared by 1) dissolving at least one compound of Formulas (I), (II),(III), (IV), or (V) in an oily phase, such as, for example, a mixture ofsoybean oil and lecithin; 2) combining the Formulas (I), (II), (III),(IV), or (V) containing oil phase with a water and glycerol mixture; and3) processing the combination to form a microemulsion.

A sterile aqueous or oleaginous suspension can be prepared in accordancewith methods already known in the art. For example, a sterile aqueoussolution or suspension can be prepared with a non-toxicparenterally-acceptable diluent or solvent, such as, for example,1,3-butane diol; and a sterile oleaginous suspension can be preparedwith a sterile non-toxic acceptable solvent or suspending medium, suchas, for example, sterile fixed oils, e.g., synthetic mono- ordiglycerides; and fatty acids, such as, for example, oleic acid.

Pharmaceutically acceptable carriers, adjuvants, and vehicles that maybe used in the pharmaceutical compositions of this invention include,but are not limited to, ion exchangers, alumina, aluminum stearate,lecithin, self-emulsifying drug delivery systems (SEDDS) such asd-alpha-tocopherol polyethyleneglycol 1000 succinate, surfactants usedin pharmaceutical dosage forms such as Tweens, polyethoxylated castoroil such as CREMOPHOR® surfactant (BASF), or other similar polymericdelivery matrices, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, potassium sorbate,partial glyceride mixtures of saturated vegetable fatty acids, water,salts or electrolytes, such as protamine sulfate, disodium hydrogenphosphate, potassium hydrogen phosphate, sodium chloride, zinc salts,colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat. Cyclodextrins such as alpha-, beta-, and gamma-cyclodextrin,or chemically modified derivatives such as hydroxyalkylcyclodextrins,including 2- and 3-hydroxypropyl-cyclodextrins, or other solubilizedderivatives may also be advantageously used to enhance delivery ofcompounds of the formulae described herein.

The pharmaceutically active compounds of this invention can be processedin accordance with conventional methods of pharmacy to produce medicinalagents for administration to patients, including humans and othermammals. The pharmaceutical compositions may be subjected toconventional pharmaceutical operations such as sterilization and/or maycontain conventional adjuvants, such as preservatives, stabilizers,wetting agents, emulsifiers, buffers etc. Tablets and pills canadditionally be prepared with enteric coatings. Such compositions mayalso comprise adjuvants, such as wetting, sweetening, flavoring, andperfuming agents.

The amounts of compounds that are administered and the dosage regimenfor treating a disease condition with the compounds and/or compositionsof this invention depends on a variety of factors, including the age,weight, sex, the medical condition of the subject, the type of disease,the severity of the disease, the route and frequency of administration,and the particular compound employed. Thus, the dosage regimen may varywidely, but can be determined routinely using standard methods. A dailydose of about 0.001 to 100 mg/kg body weight, preferably between about0.0025 and about 50 mg/kg body weight and most preferably between about0.005 to 10 mg/kg body weight, may be appropriate. The daily dose can beadministered in one to four doses per day. Other dosing schedulesinclude one dose per week and one dose per two day cycle.

For therapeutic purposes, the active compounds of this invention areordinarily combined with one or more adjuvants appropriate to theindicated route of administration. If administered orally, the compoundsmay be admixed with lactose, sucrose, starch powder, cellulose esters ofalkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesiumstearate, magnesium oxide, sodium and calcium salts of phosphoric andsulfuric acids, gelatin, acacia gum, sodium alginate,polyvinylpyrrolidone, and/or polyvinyl alcohol, and then tableted orencapsulated for convenient administration. Such capsules or tablets maycontain a controlled-release formulation as may be provided in adispersion of active compound in hydroxypropylmethyl cellulose.

Pharmaceutical compositions of this invention comprise at least onecompound of Formulas (I), (II), (III), (IV), or (V) and/or at least onepharmaceutically acceptable salt thereof, and optionally an additionalagent selected from any pharmaceutically acceptable carrier, adjuvant,and vehicle. Alternate compositions of this invention comprise acompound of the Formulas (I), (II), (III), (IV), or (V) describedherein, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

Utility

The human immune system has evolved to defend the body frommicro-organisms, viruses, and parasites that can cause infection,disease or death. Complex regulatory mechanisms ensure that the variouscellular components of the immune system target the foreign substancesor organisms, while not causing permanent or significant damage to theindividual. While the initiating events are not well understood at thistime, in autoimmune disease states the immune system directs itsinflammatory response to target organs in the afflicted individual.Different autoimmune diseases are typically characterized by thepredominate or initial target organ or tissues affected; such as thejoint in the case of rheumatoid arthritis, the thyroid gland in the caseof Hashimoto's thyroiditis, the central nervous system in the case ofmultiple sclerosis, the pancreas in the case of type I diabetes, and thebowel in the case of inflammatory bowel disease. Thus it has beenobserved that therapeutic agents which act on the immune system orcertain cell types of the immune system (such as B-lymphocytes, and Tlymphocytes, T cells) may have utility in more than one autoimmunedisease.

It is well recognized in the art, including the literature referencescited herein, that S1P receptors are good targets for a wide variety oftherapeutic applications, including autoimmune diseases. S1P receptorsmake good drug targets, because individual receptors are both tissue-and response-specific. Tissue specificity of the S1P receptors isimportant, because development of an agonist or antagonist selective forone receptor localizes the cellular response to tissues containing thatreceptor, limiting unwanted side effects. Response specificity of theS1P receptors is also important because it allows for development ofagonists or antagonists that initiate or suppress certain cellularresponses without affecting other processes. Therefore, compounds thatact on some S1P receptor family members while having diminished or noactivity at other family members are desirable and are expected toprovide a therapeutic effect with an improved side effect profile (i.e.,reduction or elimination of unwanted side effects).

As used herein, the term “agonist” in reference to S1P₁ refers to anagent which exerts pharmacological effects such as decreased motility ofT cells, decreased trafficking of T cells, or decreased egress of Tcells from lymphoid tissues. (Rosen et al., Trends in Immunology, 28:102(2007)).

By virtue of their S1P₁ activity as agonists, the compounds of thepresent invention are immunoregulatory agents useful for treating orpreventing autoimmune or chronic inflammatory diseases. The compounds ofthe present invention are useful to suppress the immune system ininstances where immunosuppression is in order, such as in bone marrow,organ or transplant rejection, autoimmune and chronic inflammatorydiseases, including systemic lupus erythematosis, rheumatoid arthritis,type I diabetes mellitus, inflammatory bowel disease, biliary cirrhosis,uveitis, multiple sclerosis, Crohn's disease, ulcerative colitis,bullous pemphigoid, sarcoidosis, psoriasis, autoimmune myositis,Wegener's granulomatosis, ichthyosis, Graves ophthalmopathy, and asthma.

More particularly, the compounds of the present invention are useful totreat or prevent a disease or disorder selected from the groupconsisting of: transplantation of organs or tissue, graft-versus-hostdiseases brought about by transplantation, autoimmune syndromesincluding rheumatoid arthritis, juvenile idiopathic arthritis, systemiclupus erythematosus, cutaneous lupus erythematosus (discoid lupuserythematosus, subacute lupus erythematosus) and lupus nephritis,Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type Idiabetes, uveitis, posterior uveitis, allergic encephalomyelitis,glomerulonephritis, post-infectious autoimmune diseases includingrheumatic fever and post-infectious glomerulonephritis, inflammatory andhyperproliferative skin diseases, psoriasis, psoriatic arthritis, atopicdermatitis, contact dermatitis, eczematous dermatitis, seborrhoeicdermatitis, lichen planus, pemphigus, bullous pemphigoid, epidermolysisbullosa, urticaria, angioedemas, vasculitis including ANCA-associatedvasculitis, giant cell arteritis, Takayasu's arteritis, microscopicpoliangiitis, central nervous system vasculitis, Churg-Strauss Syndrome,and rheumatoid vasculitis, erythema, cutaneous eosinophilia, acne,alopecia areata, keratoconjunctivitis, vernal conjunctivitis, uveitisassociated with Behcet's disease, keratitis, herpetic keratitis, conicalcornea, dystrophia epithelialis corneae, corneal leukoma, ocularpemphigus, Mooren's ulcer, scleritis, Graves' opthalmopathy,Vogt-Koyanagi-Harada syndrome, sarcoidosis, pollen allergies, reversibleobstructive airway disease, bronchial asthma, allergic asthma, intrinsicasthma, extrinsic asthma, dust asthma, chronic or inveterate asthma,late asthma and airway hyper-responsiveness, bronchitis, gastric ulcers,vascular damage caused by ischemic diseases and thrombosis, ischemicbowel diseases, inflammatory bowel diseases, necrotizing enterocolitis,intestinal lesions associated with thermal burns, coeliac diseases,proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease,ulcerative colitis, migraine, rhinitis, eczema, interstitial nephritis,Goodpasture's syndrome, hemolytic-uremic syndrome, diabetic nephropathy,multiple myositis, Guillain-Barre syndrome, Meniere's disease,polyneuritis, multiple neuritis, mononeuritis, radiculopathy,hyperthyroidism, Basedow's disease, pure red cell aplasia, aplasticanemia, hypoplastic anemia, idiopathic thrombocytopenic purpura,autoimmune hemolytic anemia, agranulocytosis, pernicious anemia,megaloblastic anemia, anerythroplasia, osteoporosis, sarcoidosis,fibroid lung, idiopathic interstitial pneumonia, dermatomyositis,leukoderma vulgaris, ichthyosis vulgaris, photoallergic sensitivity,cutaneous T cell lymphoma, arteriosclerosis, atherosclerosis, aortitissyndrome, polyarteritis nodosa, myocardosis, scleroderma, Wegener'sgranuloma, Sjögren's syndrome, adiposis, eosinophilic fascitis, lesionsof gingiva, periodontium, alveolar bone, substantia ossea dentis,glomerulonephritis, male pattern alopecia or alopecia senilis bypreventing epilation or providing hair germination and/or promoting hairgeneration and hair growth, muscular dystrophy, pyoderma and Sezary'ssyndrome, Addison's disease, ischemia-reperfusion injury of organs whichoccurs upon preservation, transplantation or ischemic disease,endotoxin-shock, pseudomembranous colitis, colitis caused by drug orradiation, ischemic acute renal insufficiency, chronic renalinsufficiency, toxinosis caused by lung-oxygen or drugs, lung cancer,pulmonary emphysema, cataracta, siderosis, retinitis pigmentosa, senilemacular degeneration, vitreal scarring, corneal alkali burn, dermatitiserythema multiforme, linear IgA ballous dermatitis and cementdermatitis, gingivitis, periodontitis, sepsis, pancreatitis, diseasescaused by environmental pollution, aging, carcinogenesis, metastasis ofcarcinoma and hypobaropathy, disease caused by histamine orleukotriene-C₄ release, Behcet's disease, autoimmune hepatitis, primarybiliary cirrhosis, sclerosing cholangitis, partial liver resection,acute liver necrosis, necrosis caused by toxin, viral hepatitis, shock,or anoxia, B-virus hepatitis, non-A/non-B hepatitis, cirrhosis,alcoholic cirrhosis, hepatic failure, fulminant hepatic failure,late-onset hepatic failure, “acute-on-chronic” liver failure,augmentation of chemotherapeutic effect, cytomegalovirus infection, HCMVinfection, AIDS, cancer, senile dementia, trauma, neuropathic pain,chronic bacterial infection, thrombocytopenia, IgA nephropathy,mesangioproliferative glomerulonephritis, IgG4-related disease,ankylosing spondylitis, and relapsing polychondritis. Juvenileidiopathic arthritis includes oligoarthritis-onset juvenile idiopathicarthritis, polyarthritis-onset juvenile idiopathic arthritis,systemic-onset juvenile idiopathic arthritis, juvenile psoriaticarthritis, and enthesitis-related juvenile idiopathic arthritis.

One embodiment provides a method for treating autoimmune and/orinflammatory diseases, comprising administering to a mammal in needthereof at least one compound of Formulas (I), (II), (III), (IV), or (V)or a pharmaceutically acceptable salt thereof. Another embodimentprovides the compounds of Formulas (I), (II), (III), (IV), or (V) orpharmaceutically acceptable salts thereof, for use in therapy for thetreatment of autoimmune and/or inflammatory diseases. In anotherembodiment, provided is the use of the compounds of Formulas (I), (II),(III), (IV), or (V) or pharmaceutically acceptable salts thereof, forthe manufacture of a medicament for the treatment or prophylaxis ofautoimmune and/or inflammatory disease. A therapeutically effectiveamount may be employed in these embodiments. Preferably, in theseembodiments, the autoimmune and inflammatory diseases are selected frommultiple sclerosis, rheumatoid arthritis, inflammatory bowel disease(including Crohn's disease and ulcerative colitis), psoriasis, and as anagent to prevent the rejection of transplanted organs. The method of thepresent embodiment includes administration of a therapeutically effectamount of a compound of Formulas (I), (II), (III), (IV), or (V) or apharmaceutically effective salt thereof.

In another embodiment, a method for treating vascular disease isprovided comprising administering to a mammal in need thereof at leastone compound of Formulas (I), (II), (III), (IV), or (V) or apharmaceutically acceptable salt thereof. Another embodiment providesthe compounds of Formulas (I), (II), (III), (IV), or (V) orpharmaceutically acceptable salts thereof, for use in therapy for thetreatment of vascular disease. In another embodiment, provided is theuse of the compounds of Formulas (I), (II), (III), (IV), or (V) orpharmaceutically acceptable salts thereof, for the manufacture of amedicament for treatment of vascular disease. A therapeuticallyeffective amount may be employed in these embodiments. Preferably, inthese embodiments, the vascular disease is selected from atherosclerosisand ischemia reperfusion injury.

In another embodiment, a method for treating inflammatory bowel diseaseis provided comprising administering to a mammal in need thereof atleast one compound of Formulas (I), (II), (III), (IV), or (V) or apharmaceutically acceptable salt thereof. Another embodiment providesthe compounds of Formulas (I), (II), (III), (IV), or (V) orpharmaceutically acceptable salts thereof, for use in therapy for thetreatment of inflammatory bowel disease. In another embodiment, providedis the use of the compounds of Formulas (I), (II), (III), (IV), or (V)or pharmaceutically acceptable salts thereof, for the manufacture of amedicament for treatment of inflammatory bowel disease. Atherapeutically effective amount may be employed in these embodiments.Preferably, in these embodiments, the inflammatory bowel disease isselected from Crohn's disease, ulcerative colitis, collagenous colitis,lymphocytic colitis, ischaemic colitis, diversion colitis, Behçet'sdisease, and indeterminate colitis.

In another embodiment, a method for treating lupus is providedcomprising administering to a mammal in need thereof at least onecompound of Formulas (I), (II), (III), (IV), or (V) or apharmaceutically acceptable salt thereof. Another embodiment providesthe compounds of Formulas (I), (II), (III), (IV), or (V) orpharmaceutically acceptable salts thereof, for use in therapy for thetreatment of lupus. In another embodiment, provided is the use of thecompounds of Formulas (I), (II), (III), (IV), or (V) or pharmaceuticallyacceptable salts thereof, for the manufacture of a medicament fortreatment of lupus. A therapeutically effective amount may be employedin these embodiments. Lupus includes systemic lupus erythematosus,cutaneous lupus erythematosus, discoid lupus erythematosus, subacutelupus erythematosus and lupus nephritis.

In another embodiment, a method for treating multiple sclerosis isprovided comprising administering to a mammal in need thereof at leastone compound of Formulas (I), (II), (III), (IV), or (V) or apharmaceutically acceptable salt thereof. Another embodiment providesthe compounds of Formulas (I), (II), (III), (IV), or (V) orpharmaceutically acceptable salts thereof, for use in therapy for thetreatment of multiple sclerosis. In another embodiment, provided is theuse of the compounds of Formulas (I), (II), (III), (IV), or (V) orpharmaceutically acceptable salts thereof, for the manufacture of amedicament for treatment of multiple sclerosis. A therapeuticallyeffective amount may be employed in these embodiments. Preferably, inthese embodiments, multiple sclerosis includes relapsing remittingmultiple sclerosis, primary progressive multiple sclerosis, secondaryprogressive multiple sclerosis, and progressive relapsing multiplesclerosis.

The methods of treating S1P1-associated conditions may compriseadministering compounds of Formulas (I), (II), (III), (IV), or (V) aloneor in combination with each other and/or other suitable therapeuticagents useful in treating such conditions. Accordingly, “therapeuticallyeffective amount” is also intended to include an amount of thecombination of compounds claimed that is effective to act as an agonistat the S1P1 receptor. The combination of compounds is preferably asynergistic combination. Synergy, as described, for example, by Chou etal., Adv. Enzyme Regul., 22:27-55 (1984), occurs when the effect of thecompounds when administered in combination is greater than the additiveeffect of the compounds when administered alone as a single agent. Ingeneral, a synergistic effect is most clearly demonstrated atsub-optimal concentrations of the compounds. Synergy can be in terms oflower cytotoxicity, increased efficacy, or some other beneficial effectof the combination compared with the individual components.

Exemplary of such other therapeutic agents include corticosteroids orglucocorticoids such as dexamethasone, methylprednisolone, prednisolone,and prednisone; PDE4 inhibitors such as rolipram, cilomilast,roflumilast, and oglemilast; cytokine-suppressive anti-inflammatorydrugs (CSAIDs) and inhibitors of p38 kinase, 4-substituted imidazo[1,2-A]quinoxalines as disclosed in U.S. Pat. No. 4,200,750; antibodiesor fusion proteins directed to cell surface molecules such as CD2, CD3,CD4, CD8, CD20 such as RITUXAN®, CD25, CD30, CD40, CD69, CD80 (B7.1),CD86 (B7.2), CD90, CTLA, for example abatacept (ORENCIA®), belatacept,or their ligands including CD154 (GP39, or CD40L); antibodies to, fusionproteins, or soluble receptors of human cytokines or growth factors, forexample, TNF such as, infliximab (REMICADE®), etanercept (Embrel),adalimumab (HUMIRA®), LT, Il-1 such as anakinra (KINERET®) (an IL-1receptor antagonist), IL-2, IL-4, IL-5, Il-6, such as CNTO 328 (achimeric anti-IL-6 antibody), Il-7, Il-8, Il-12, Il-15, Il-16, Il-17,Il-21, Il-23 such as Ustekinumab (a human anti-IL-12/23 monoclonalantibody), and interferons such as interferon beta 1a (AVONEX®, REBIF®),interferon beta 1b (BETASERON®); integrin receptor antagonists such asTYSABRI®; polymeric agents such as glatiramer acetate (COPAXONE®);sulfasalazine, mesalamine, hydroxychloroquine, non-steroidalantiinflammatory drugs (NSAIDs) such as salicylates including aspirin,salsalate, and magnesium salicylate, and non-salicylates such as,ibuprofen, naproxen, meloxicam, celecoxib and rofecoxib; antiviralagents such as abacavir; antiproliferative agents such as methotrexate,mercaptopurine, leflunomide, cyclosporine, mycophenololate, FK506(tacrolimus, PROGRAF®); cytotoxic drugs such as azathioprine andcyclophosphamide; nuclear translocation inhibitors, such asdeoxyspergualin (DSG); gold containing products such as auronofin;penicllamine, and rapamycin (sirolimus or RAPAMUNE®) or derivativesthereof.

The above other therapeutic agents, when employed in combination withthe compounds of the present invention, may be used, for example, inthose amounts indicated in the Physicians' Desk Reference (PDR) or asotherwise determined by one of ordinary skill in the art. In the methodsof the present invention, such other therapeutic agent(s) may beadministered prior to, simultaneously with, or following theadministration of the inventive compounds.

Methods of Preparation

The compounds of the present invention can be prepared in a number ofways well known to one skilled in the art of organic synthesis. Thecompounds of the present invention can be synthesized using the methodsdescribed below, together with synthetic methods known in the art ofsynthetic organic chemistry, or variations thereon as appreciated bythose skilled in the art. Preferred methods include, but are not limitedto, those described below. All references cited herein are herebyincorporated in their entirety by reference.

The compounds of this invention may be prepared using the reactions andtechniques described in this section. The reactions are performed insolvents appropriate to the reagents and materials employed and aresuitable for the transformations being effected. Also, in thedescription of the synthetic methods described below, it is to beunderstood that all proposed reaction conditions, including choice ofsolvent, reaction atmosphere, reaction temperature, duration of theexperiment and work up procedures, are chosen to be the conditionsstandard for that reaction, which should be readily recognized by oneskilled in the art. It is understood by one skilled in the art oforganic synthesis that the functionality present on various portions ofthe molecule must be compatible with the reagents and reactionsproposed. Such restrictions to the substituents that are compatible withthe reaction conditions will be readily apparent to one skilled in theart and alternate methods must then be used. This will sometimes requirea judgment to modify the order of the synthetic steps or to select oneparticular process scheme over another in order to obtain a desiredcompound of the invention. It will also be recognized that another majorconsideration in the planning of any synthetic route in this field isthe judicious choice of the protecting group used for protection of thereactive functional groups present in the compounds described in thisinvention. An authoritative account describing the many alternatives tothe trained practitioner is Greene and Wuts (Protective Groups InOrganic Synthesis, Fourth Edition, Wiley and Sons, 2006).

Compounds of Formula (I) may be prepared by reference to the methodsillustrated in the following Schemes. As shown therein the end productis a compound having the same structural formula as Formula (I). It willbe understood that any compound of Formula (I) may be produced by theschemes by the suitable selection of reagents with appropriatesubstitution. Solvents, temperatures, pressures, and other reactionconditions may readily be selected by one of ordinary skill in the art.Starting materials are commercially available or readily prepared by oneof ordinary skill in the art. Constituents of compounds are as definedherein or elsewhere in the specification.

As shown in Scheme 1, compounds of Formula I may be produced, startingwith bicyclic compounds 1.1 in which an aryl or heteroaryl boronic acidcan be coupled with cyclopentenone in a conjugate addition reaction(catalyzed for example with rhodium or copper complexes) to affordketone 1.2. This transformation can be done in the presence of chiralligands (such as BINAP) to provide enantioenriched 1.2. Ketone 1.2 canalso be prepared through transition-metal catalyzed coupling of aryl orheteroaryl halogen compounds with cyclopeten-2-enol. Ketone 1.2 can beconverted to either amino-nitrile 1.3 or hydantoin 1.4, each of whichcan be hydrolyzed to provide amino-acid 1.5. Direct reduction of theacid of 1.5, or initial esterification and subsequent reduction of thecarbonyl ester, leads to compounds of formula I.

Alternatively, I can be obtained from 2.3 through reduction of theolefin and carbonyl. 2.3 can be prepared through the transition-metalmediated coupling of 1.1 and 2.2, with 2.2 coming from the coupling of2.1 and 1,4-dichlorobut-2-ene under basic conditions.

Compounds of formula I can be prepared from carbonyl compounds 3.1 asshown in Scheme 3 by reduction to alcohols 3.2 followed by alkylation toafford ethers 3.5. Alternatively, condensation of 3.1 with alcoholsaffords ketals 3.3 or enol ethers 3.4, either of which can be reactedunder reducing conditions (such as palladium catalyzed hydrogenation) toprovide ethers 3.5. Conversion of 3.1 to enol triflate 3.6 can befollowed by metal-mediated coupling to afford alkyl, aryl, or heteroarylderivatives 3.7.

Enol triflate 3.6 can also be converted to carbonyl ester derivatives4.1 which can be further reduced to alcohols 4.2. Alkylation of 4.2 canprovide ethers 4.3 while conversion of the alcohol to a leaving group(e.g. halogen or tosylate) followed by displacement with a nucleophilecan lead to 4.3 as ether, amine or thioether derivatives. Oxidation of4.2 followed by olefination leads to 4.5 which can be further reduced to4.6.

Preparation of bicyclic frameworks useful for the invention are outlinedin Scheme 5. Elaboration of 5.1 to carboxylic esters 5.2, hydrolysisunder acidic or basic conditions to provide acid 5.3, conversion to acidchloride 5.4, followed by cationic cyclization in the presence of aterminal olefin affords ketones 5.3 which can be further modified asdescribed above in Schemes 3 and 4 to for example compounds 5.6. 5.1 canalso be coupled with olefins 5.9 under palladium catalysis to afford5.10, which can undergo reduction of the alkene and then cyclizationunder acidic conditions (such as PPA or H₂SO₄) to give bicyclic 5.12.Reduction of the ketone of 5.12 affords 5.13.

EXAMPLES

The invention is further defined in the following Examples. It should beunderstood that the Examples are given by way of illustration only. Fromthe above discussion and the Examples, one skilled in the art canascertain the essential characteristics of the invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications to adapt the invention to various uses and conditions.As a result, the invention is not limited by the illustrative examplesset forth hereinbelow, but rather is defined by the claims appendedhereto.

ABBREVIATIONS

Ac acetyl

AcOH acetic acid

anhyd. anhydrous

aq. aqueous

BH₃.DMS borane-dimethyl sulfide

BF₃.Et₂O boron trifluoride diethyl etherate

Bn benzyl

BOC₂O di-tert-butyl dicarbonate

Bu butyl

Boc tert-butoxycarbonyl

CV Column Volumes

DCE dichloroethane

DCM dichloromethane

DEA diethylamine

DIEA diisopropylethylamine

DMA N,N-dimethylacetamide

DMF dimethylformamide

DMPU 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone

DMSO dimethylsulfoxide

EtOAc ethyl acetate

Et ethyl

Et₃N triethyl amine

EtOH ethanol

H or H₂ hydrogen

h, hr or hrs hour(s)

hex or Hex hexane

i iso

IPA isopropyl alcohol

HOAc acetic acid

HCl hydrochloric acid

HPLC high pressure liquid chromatography

i-PrOH isopropanol

KHMDS potassium bis(trimethylsilyl) amide

LC liquid chromatography

LCMS liquid chromatography mass spectroscopy

LDA lithium diisopropylamine

LiHMDS lithium bis(trimethylsilyl) amide

m-CPBA meta-chloroperoxybenzoic acid

M molar

mM millimolar

Me methyl

MeCN acetonitrile

Mel methyl iodide

MeOH methanol

MHz megahertz

min. minute(s)

mins minute(s)

M⁺¹ (M+H)⁺

MS mass spectrometry

n or N normal

NIS N-Iodosuccinimide

nm nanometer

nM nanomolar

NMO N-methylmorpholine-N-oxide

NMP N-methylpyrrolidine

Pd/C palladium on carbon

Pd(OAc)₂ palladium acetate

Pd(PPh₃)₄ tetrakis(triphenylphosphine)palladium

Pd₂(dba)₃ tris-(dibenzylideneacetone)dipalladium

Ph phenyl

PPA polyphosphoric acid

PPC pyrophosphoryl chloride

PPh₃ triphenylphosphine

Pr propyl

PSI pounds per square inch

Ret Time or Rt retention time

sat. saturated

S-BINAP S)-(−)-2,2′-bis(diphenylphosphino)-1,1′-binaphthyl

SFC supercritical fluid chromatography

t-BuOH tertiary butanol

TFA trifluoroacetic acid

THF tetrahydrofuran

Analytical HPLC Conditions:

Condition A: Column: Waters Acquity UPLC BEH C18, 2.1×50 mm, 1.7-μmparticles; Mobile Phase A: 5:95 acetonitrile:water with 0.05% TFA;Mobile Phase B: 95:5 acetonitrile:water with 0.05% TFA; Temperature: 50°C.; Gradient: 0-100% B over 3 minutes, then a 0.75-minute hold at 100%B; Flow: 1.11 mL/min.Condition B: Column: 1-Waters C18 2.1×30 mm 3.5 um (4 min.); SolventA=10% MeOH, 90% H₂O, 0.1% TFA; Solvent B=90% MeOH, 10% H₂O, 0.1% TFA.Condition C: Column: YMC CombiScreen S5 50×4.6 mm (4 min; SolventA=Water 90%/MeOH 10%/H₃PO₄, 0.2%; Solvent B=MeOH 90%/water 10%/H₃PO₄0.2%.Condition G: Column: Waters Acquity BEH C18 2.1×50 mm 1.7 um; Lineargradient of 0-100% solvent B over 3 min, then 0.75 min hold at 100% B;Flow rate: 1.11 mL/min; Solvent A: 5:95 acetonitrile:water with 10 mMammonium acetate; Solvent B: 95:5 acetonitrile:water with 10 mM ammoniumacetate; Temperature=50° C.; Products detected at 220 nm wavelength w/positive ionization mode.Condition H: Column: Sunfire C18, (150×3.0 mm), 3.5 pm; Linear gradientof 10 to 100% solvent B over 25 min, then 5 min hold at 100% B; Flowrate: 1 mL/min; Buffer: 0.5% TFA, in water with pH adjusted to 2.5 usingdilute ammonia; Solvent A: Buffer: acetonitrile (95:5); Solvent B:Buffer: acetonitrile (5:95); Products detected at 220 nm.Condition I: Column: Xbridge Phenyl, (150×3.0 mm), 3.5 pm; Lineargradient of 10 to 100% solvent B over 25 min, then 5 min hold at 100% B;Flow rate: 1 mL/min; Buffer: 0.5% TFA, in water with pH adjusted to 2.5using dilute ammonia; Solvent A: Buffer: acetonitrile (95:5); Solvent B:Buffer: acetonitrile (5:95); Products detected at 220 nm.Condition J: Column: Chromolith SpeedROD (4.6×50 mm); Linear gradient of0 to 100% solvent B over 4 min, with 1 min hold at 100% B; Solvent A:10% MeOH, 90% H₂O, 0.1% TFA; Solvent B: 90% MeOH, 10% H₂O, 0.1% TFA;Flow rate: 4 mL/min; Products detected at 220 nm.Condition K: Column: YMC ProC18 S5 ODS (50×4.6 mm); Linear gradient of 0to 100% solvent B over 4 min, with 1 min hold at 100% B Solvent A: 10%MeOH-90% H₂O-0.2% H₃PO₄; Solvent B: 90% MeOH-10% H₂O-0.2% H₃PO₄; Flowrate: 4 mL/min; Products detected at 220 nm.Condition L: Column: Sunfire C18 3.5 um, 3.0×150 mm; Linear gradient of10 to 100% solvent B over 12 min, with 3 min hold at 100% B; SolventA=0.05% TFA in H₂O:MeCN (95:5); Solvent B=0.05% TFA in H₂O:MeCN (5:95).Flow rate: 1 mL/min; Products detected at 220 nm and 256 nm.Condition M: Waters Acquity BEH C18 2.1×50 mm 1.7 um; Linear gradient of0-100% solvent B over 1.5 min 100% B; Flow rate: 1 mL/min; Solvent A:10:90 acetonitrile:water with 0.1% TFA; Solvent B: 90:10acetonitrile:water with 0.1% TFA; Temperature=40° C.; Products detectedat 220 nm wavelength w/ positive ionization mode.Condition Gemini:Column:Phenomenex Gemini C18, 3 μm, 4.6×150 mm; Grad.T: 10 min; Flow R.: 1.0 mL/min.; Solvent Grad.: 30-100% B; Wave: 220 nm.(A=5% MeCN-90% H₂O-0.1% TFA; B=95% MeCN-5% H₂O-0.1% TFA).

EXAMPLES

The following examples illustrate the particular and preferredembodiments of the present invention and do not limit the scope of thepresent invention. Chemical abbreviations and symbols as well asscientific abbreviations and symbols have their usual and customarymeanings unless otherwise specified. Additional abbreviations employedin the Examples and elsewhere in this application are defined above.Common intermediates are generally useful for the preparation of morethan one Example and are identified sequentially (e.g., Intermediate 1,Intermediate 2, etc. and are abbreviated as Int. 1, Int. 2, etc.Compounds of the Examples are identified by the example and step inwhich they were prepared (e.g., “1-A” denotes the Example 1, step A), orby the example only where the compound was the title compound of theexample (for example, “1” denotes the title compound of Example 1). Insome instances alternate preparations of intermediates or examples aredescribed. Frequently chemists skilled in the art of synthesis maydevise alternative preparations which may be desirable based on one ormore considerations such as shorter reaction time, less expensivestarting materials, ease of operation, amenable to catalysis, avoidanceof toxic reagents, accessibility of specialized instrumentation, anddecreased number of linear steps, etc. The intent of describingalternative preparations was to further enable the preparation of theexamples of this invention. In some instances some functional groups inthe outlined examples and claims may be replaced by well known biostericreplacements known in the art, for example, replacement of a carboxylicacid group with a tetrazole or a phosphate moiety.

Those experiments specifying that they were performed in a microwaveoven were conducted in a SmithSynthesizer™ oven manufactured by PersonalChemistry or a Discover™ microwave oven manufactured by CEM corporation.The microwave ovens generate a temperature which can be selected to bebetween 60-250° C. The microwave ovens automatically monitor thepressure which was between 0-300 PSI. Reaction hold times andtemperature set points are reported.

Intermediate 1 (1R,3S)-methyl1-amino-3-(4-bromophenyl)cyclopentanecarboxylate

Intermediate 1A: (S)-3-(4-bromophenyl)cyclopentanone

A solution of 4-bromophenylboronic acid (20 g, 100 mmol) in 1,4-dioxane(120 mL) in a 500 ml flask was purged with nitrogen for 5 mins. S-BINAP(0.992 g, 1.593 mmol) and bis(norbornadiene)rhodium (I)tetrafluoroborate (0.559 g, 1.494 mmol) were added sequentially to thesolution under a positive pressure of nitrogen. After 2 hours ofagitation at room temperature, water (20 mL) was added followed bycyclopent-2-enone (8.06 mL, 100 mmol) and Et₃N (13.88 mL, 100 mmol). Themixture was allowed to stir at room temperature for 16 hours. Theresulting dark solids were removed by filtration and the filtrate waspoured into 250 ml of ethyl acetate. The solution was washed with watertwice and the organic layer was concentrated. The residue was purifiedby flash column chromatography (split into two batches, each run on a330 g silica column. 0%-25% ethyl acetate in hexane) to afford 12.1grams of (S)-3-(4-bromophenyl) cyclopentanone. HPLC purity was >98% andChiral HPLC analysis indicated approximately 90% ee. The material wasfurther purified by under the Chiral SFC using the following conditions:Instrument: Berger SFC MGIII; Preparation Conditions: Column: ChiralPakAD-H 25×5 cm, 5 μm; Column Temp. 40° C.; Flow rate: 200 ml/min; MobilePhase: CO₂/MeOH=80/20; Detector Wavelength: 225 nm; AnalyticalConditions Injection Vol. 1.0 ml; Sample Preparation: 12.1 g in 210 mLMeOH (Conc. 60 mg/ml); Column: ChiralPak AD 25×0.46 cm, 10 μm; ColumnTemp. 40° C.; Flow rate: 2.0 min; Mobile Phase: CO₂/MeOH=70/30; DetectorWavelength: 220 nm; Injection Vol. 5 μL.

The desired enantiomer (major isomer) was isolated and named as “PK2”based on the elution order. The enantiomeric purity of the isolatedisomer was determined to be greater than 99.6% on SFC/UV area % at 220nm. After concentration, 10.5 grams of the desired enantiomer wasrecovered. HPLC retention time=8.19 min (condition G); LC/MS M⁺¹=240.08;¹H NMR ((400 MHz, CD₃OD) δ ppm 7.43-7.51 (2H, m), 7.10-7.19 (2H, m),3.32-3.46 (1H, m), 2.67 (1H, dd, J=18.27, 7.48 Hz), 2.39-2.54 (2H, m),2.23-2.39 (2H, m), 1.97 (1H, ddd, J=12.98, 11.00, 9.02 Hz).

Intermediate 1B:(7S)-7-(4-bromophenyl)-1,3-diazaspiro[4.4]nonane-2,4-dione

A total of 9.8 g (S)-3-(4-bromophenyl)cyclopentanone was used, dividedinto two batches each containing 4.9 g. The two batches were processedunder identical conditions as described below.

To a mixture of (S)-3-(4-bromophenyl)cyclopentanone (4.9 g, 20.49 mmol)and potassium cyanide (1.935 g, 29.7 mmol) in EtOH (40 mL) and water (20mL) in a glass pressure vessel was added ammonium carbonate (4.92 g,51.2 mmol). The reaction vessel was sealed and placed in an oil bathheated at 80° C. for 24 hours, resulting in the formation of a whitesolid. After cooling the reaction vessel with an ice bath ice-bath, thevessel was opened and 30 ml of water was add resulting in the formationof additional solids. The solids were collected by filtration, washedtwice with 5 ml water, then dried under high vacuum. The two batcheswere combined (total 13.9 g(7S)-7-(4-bromophenyl)-1,3-diazaspiro[4.4]nonane-2,4-dione) and thematerial was used without further purification for subsequent steps.HPLC retention time=0.82 min (condition G) LC/MS M⁺¹=331.1. ¹H NMR (400MHz, MeOD) δ ppm 7.43 (2H, d, J=7.7 Hz), 7.22 (2H, dd, J=8.4, 6.2 Hz),2.31-2.43 (1H, m), 2.17 (3H, d, J=9.9 Hz), 1.79-2.06 (3H, m).

Intermediate 1C: (3S)-1-amino-3-(4-bromophenyl)cyclopentanecarboxylicAcid

To (7S)-7-(4-bromophenyl)-1,3-diazaspiro[4.4]nonane-2,4-dione (13.9 g,45.0 mmol) in 1,4-dioxane (40 mL) in a round bottom flask was addedaqueous NaOH (2N, 100 mL, 200 mmol). The mixtures were heated to 95° C.and stirred for 24 hours. Additional NaOH (25 mL, 50 mmol) was added andheating was continued for another two days. The solution was cooled withan ice-bath, neutralized with 5N HCl to approximately pH 7 resulting inthe formation of a white precipitate. The solids were collected byfiltration and dried under high vacuum for 2 days to provide 14 g of(3S)-1-amino-3-(4-bromophenyl)cyclopentanecarboxylic acid as whitesolid. The material was used directly in the subsequent step withoutfurther purification. HPLC retention time=0.64 min (condition G) LC/MSM⁺¹=284.1/286.1.

Intermediate 1D: (3S)-methyl1-amino-3-(4-bromophenyl)cyclopentanecarboxylate

To a heterogeneous mixture of (3S)-1-amino-3-(4-bromophenyl)cyclopentanecarboxylic acid (14 g, 49.3 mmol) in MeOH (250 mL) was addedthionyl chloride (36.0 mL, 493 mmol) dropwise over a period of 20 min.at room temperature via an additional funnel (exothermic). The reactionmixture was placed in an oil bath set to 70° C. for 4 hours. The solventwas removed under vacuum, with the residue being dissolved in ethylacetate (200 mL) and washed twice with 1N NaOH. The organic layer wasthen dried over Na₂SO₄ and concentrated to give 10.8 g of (3S)-methyl1-amino-3-(4-bromophenyl)cyclopentanecarboxylate. HPLC retentiontime=0.68 min (condition G); LC/MS M⁺¹=298/300.

Intermediate 1: (1R,3S)-methyl1-amino-3-(4-bromophenyl)cyclopentanecarboxylate

The mixture of diastereomers (I-1D, 9.5 g) was separated by Chiral SFC.The absolute stereochemical assignment of Intermediate 1 and itsdiastereomer was previously described (Wallace, G. A. et al. J. Org.Chem. 2009, 74, 4886-4889). Experimental Details: Instrument:Preparative: Thar SFC350; Analytical: Berger analytical SFC; PreparativeConditions: Column: Lux-Cellulose-4 25×3 cm, 5 μm; Column Temperature:35° C.; Flow rate: 200 ml/min; Mobile Phase: CO₂/(MeOH with 0.1%DEA)=87/13; Detector Wavelength: 220 nm; Injection Vol.: 0.6 ml; SamplePreparation: 9.5 g in 400 ml MeOH (Conc. 23.7 mg/ml). AnalyticalConditions: Column: Lux-Cellulose-4 25×0.46 cm, 5 μm; Column Temp. 35°C.; Flow rate: 3 ml/min; Mobile Phase: CO₂/(MeOH with 0.1% DEA)=85/15;Detector Wavelength: 220 nm; Injection Vol.: 5 μL. Intermediate 1 wasPeak 2: 4.06 g; ret. time=6.64 min on the analytical chiral SFCconditions above. Optical purity: 98.2%; LC/MS M⁺¹=298/300; Peak 1: 3.96g; ret. time=5.47 min on the analytical chiral SFC conditions above.Optical purity: 99.4%. ¹H NMR (400 MHz, DMSO-d₆) δ 8.89 (br. s., 2H),7.51 (d, J=8.1 Hz, 2H), 7.34 (d, J=8.4 Hz, 2H), 3.80 (s, 3H), 2.59 (dd,J=13.6, 7.5 Hz, 2H), 2.30-1.94 (m, 5H). Peak 1: 3.96 g; ret. time=5.47min on the analytical chiral SFC conditions above. Optical purity:99.4%.

Alternative Preparation: HCl Salt of Intermediate 1

A solution of (3S)-1-amino-3-(4-bromophenyl)cyclopentanecarboxylic acid(10.2 g, 35.9 mmol) in MeOH (100 mL) was cooled in an ice bath, followedby addition of SOCl₂ (15.72 mL, 215 mmol) dropwise. After the additionwas complete, the solution was refluxed for 3 hrs at which time thereaction was determined to be complete by EA-HPLC. The solution wasconcentrated to remove the methanol to afford a solid. The solid wastaken in 50 ml of 3% H₂O in EtOAc and stirred well for 30 mins. Thewhite solid formed was collected by filtration and the wet white solidwas taken in 50 ml of 4% H₂O in 1,2-dimethoxyethane and heated to 50° C.for 3 hrs, then stirred at room temperature overnight. The resultingwhite solid was collected by filtration and dried to afford product(1R,3S)-methyl 1-amino-3-(4-bromophenyl)cyclopentanecarboxylatehydrochloride (3.5 g, 10.35 mmol). HPLC retention time=6.6 min(condition H) LC/MS M⁺¹=298/300. ¹H NMR (400 MHz, DMSO-d₆) δ 8.95 (br.s, 3H) 7.50-7.53 (m, 2H), 7.35-7.37 (m, 2H), 3.81 (s, 3H) 3.17-3.28 (m,1H), 2.57 (dd, J=14, 7 Hz, 1H), 2.0-2.28 (m, 5H).

Intermediate 2 (1R,3R)-methyl1-amino-3-(4-bromophenyl)cyclopentanecarboxylate Br OCH₃

Intermediate 2A: (R)-3-(4-bromophenyl)cyclopentanone

A solution of 4-bromophenylboronic acid (20 g, 100 mmol) in 1,4-dioxane(120 mL) was purged with nitrogen for 10 min. (R)-BINAP (0.992 g, 1.593mmol) and bis(norbornadiene)rhodium (I) tetrafluoroborate (0.559 g,1.494 mmol) were added sequentially, and the suspension was sonicatedfor 5 min. The mixture was stirred for 20 min. Water (20 mL) was added,and the reaction mixture became homogeneous. After 10 minutes,cyclopent-2-enone (8.06 mL, 100 mmol) was added, and the reactionmixture was stirred at room temperature overnight. HPLC and LCMSanalysis indicated that the reaction had proceeded, but there was morestarting material than product. The reaction mixture was filteredthrough a pad of Celite, and the Celite was washed with ethyl acetate(100 mL). The filtrate was diluted with an additional ethyl acetate (150mL), washed with water (2×), washed with brine, and dried over anhydroussodium sulfate. The product mixture was purified by flash silica gelchromatography using a mixture of ethyl acetate and hexane to give(R)-3-(4-bromophenyl)cyclopentanone (6.09 g, 25.5 mmol) as a whitesolid. The product was 98% pure by HPLC with a ret. time=2.11 min.(Condition J). LC/MS M⁺¹=241. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.57-7.39(m, 2H), 7.22-7.06 (m, 2H), 3.39 (ddd, J=10.9, 6.8, 4.1 Hz, 1H), 2.67(dd, J=18.2, 7.4 Hz, 1H), 2.57-2.38 (m, 2H), 2.38-2.21 (m, 2H),1.99-1.85 (m, 1H).

Chiral HPLC indicated that the compound was 90-95% enantiomericallypure. The compound (6.03 g) was further purified by Chiral SFC using theconditions listed below. The desired enantiomer was isolated and namedas “PK1” in the elution order. The enantiomeric purity of the isolatedisomer was determined to be greater than 99.9% on SFC/UV area % at 220nm. The desired enantiomer (5.45 g) was recovered after concentration.Experimental Details: Instrument: Berger SFC MGIII; Prep. Conditions;Column: ChiralPak AD-H 25×3 cm, 5 μm; Column Temperature: 40° C.; Flowrate: 180 ml/min; Mobile Phase: CO₂/MeOH=87/13; Detector Wavelength: 225nm; Injection Vol.: 0.5 ml; Sample Preparation: 6.03 g in 100 mL MeOH(Conc. 60 mg/ml). Analytical Conditions: Column: ChiralPak AD 25×0.46cm, 10 μm; Column Temperature: 40° C.; Flow rate: 2.0 min; Mobile Phase:CO₂/MeOH=70/30; Detector Wavelength: 220 nm; Injection Vol.: 5 μL.

Intermediate 2B:(7R)-7-(4-bromophenyl)-1,3-diazaspiro[4.4]nonane-2,4-dione

To a mixture of (R)-3-(4-bromophenyl)cyclopentanone (5.4 g, 22.58 mmol)and potassium cyanide (2.132 g, 32.7 mmol) in EtOH (40 mL) and water (20mL) in a glass pressure vessel was added ammonium carbonate (5.42 g,56.5 mmol). The reaction vessel was sealed and placed in an oil bathheated at 80° C. for 20 hours. A large amount of white, free flowingsolid formed in the pale yellow solution. Analysis by LCMS indicatedremaining starting material so the reaction was continued for anadditional 24 hours. As conversion was incomplete, the temperature ofthe oil bath was raised to 120° C. The white solid completely dissolvedat the higher temperature. After 3 hours the solution was cooled down toroom temperature. The solution was further cooled in an ice bath, water(30 mL) was added and the resulting white solid was collected byfiltration, washed with water, air dried, then placed under high vacuumto afford the target compound (6.9 g, 22.32 mmol) which was used forsubsequent reaction without additional purification. HPLC retentiontime=0.81 min (condition G); LC/MS M⁺¹=309/311; 2M^(+H)=619.

Intermediate 2C: (3R)-1-amino-3-(4-bromophenyl)cyclopentanecarboxylicAcid

A solution of (7R)-7-(4-bromophenyl)-1,3-diazaspiro[4.4]nonane-2,4-dione(6.80 g, 22 mmol) in dioxane (20 mL) and NaOH (2N aq) (120 mL, 240 mmol)was heated in an oil bath set to 95° C. The resulting clear, pale yellowsolution was left to stir over the weekend. The solution was cooled inan ice bath and neutralized to approximately pH 7 with 6 N HCl resultingin the formation of a precipitate. The solids were collected and left toair dry overnight. The white solid was slurried in hot ethanol (˜100 mL)and re-collected by filtration and the solid was air-dried then placedunder high vacuum. (5.8 g, 20.41 mmol). HPLC retention time=0.64 min(condition G); LC/MS M⁺¹=284/286. ¹H NMR (500 MHz, METHANOL-d₄) δ7.52-7.38 (m, 2H), 7.31-7.17 (m, 2H), 3.55-3.40 (m, 1H), 2.68 (dd,J=13.3, 6.7 Hz, 1H from single diastereomer), 2.58-2.39 (m, 1H),2.26-2.15 (m, 1H), 2.10-1.98 (m, 1H), 1.98-1.81 (m, 1H), 1.70 (dd,J=13.2, 11.8 Hz, 1H from single diastereomer).

Intermediate 2D: (3R)-methyl1-amino-3-(4-bromophenyl)cyclopentanecarboxylate

In a 500 mL round bottom flask containing a stir bar,(3R)-1-amino-3-(4-bromophenyl) cyclopentanecarboxylic acid (5.4 g, 19.00mmol) was suspended in methanol (100 mL) to afford a white slurry. Adropping funnel was charged with thionyl chloride (13.87 mL, 190 mmol)and the reagent was added dropwise at a rate to keep the mixture fromreaching reflux temperature. After the addition was complete, the paleyellow, milky solution was placed in an oil bath set to 70° C. and anair-cooled reflux condenser was attached. The solution was heated forseveral hours and then allowed to cool to room temperature overnight.The solvent was evaporated under vacuum. The residue was dissolved inethyl acetate, washed with 1N NaOH (aq), washed with water, then driedover MgSO₄ before being filtered and concentrated. The resulting yellowsolid was slurried in warm ethyl acetate, with sonication and thenfiltered. The solid was air-dried and placed under vacuum and thefiltrate was evaporated to afford Solid 1: white solid, 4.28 g LCMSshows >98% AP. The filtrate was evaporated to afford a yellow solid(1.89 g). The solid from the filtrate was slurried in a minimal amountof hot ethyl acetate, with sonication, then cooled (ice bath) andfiltered cold. The solid was air-dried and placed under vacuum to affordSolid 2: 1.44 g white solid. Combined solids (5.7 g).

Intermediate 2: (1R,3R)-methyl1-amino-3-(4-bromophenyl)cyclopentanecarboxylate

The combined solids of (3R)-methyl 1-amino-3-(4-bromophenyl)cyclopentanecarboxylate (4 g) were separated using Chiral SFC separationof the diastereomers. The absolute stereochemical assignment ofIntermediate 2 and its diastereomer has been previously described(Wallace, G. A. et al. J. Organic Chem. 2009, 74, 4886-4889).Experimental Details: Instrument: Preparative: Thar SFC350; Analytical:Thar analytical MDS. Preparative Conditions: Column: ChiralPak AD-H 25×5cm, 5 μm; Column Temperature: 35° C.; Flow rate: 300 ml/min; MobilePhase: CO₂/(MeOH with 0.1% DEA)=82/18; Detector Wavelength: 230 nm;Injection Vol.: 0.4-0.5 ml; Sample Preparation: 4 g in 120 ml MeOH(Conc. 33 mg/ml). Analytical Conditions: Column: ChiralPak AD-H 25×0.46cm, 5 μm; Column Temperature: 35° C.; Flow rate: 3 ml/min; Mobile Phase:CO₂/(MeOH with 0.1% DEA)=80/20; Detector Wavelength: 222 nm; InjectionVol.: 5 μL. Intermediate 2 was Peak 1: 1.56 g (99.3% optical purity at222 nm) Ret. Time=7.18 min on analytical chiral SFC. ¹H NMR (500 MHz,METHANOL-d₄) δ 7.45-7.39 (m, 2H), 7.23-7.17 (m, 2H), 3.78 (s, 3H),3.40-3.48 (m, 1H), 2.40 (ddd, J=13.0, 8.9, 3.6 Hz, 1H), 2.28-2.21 (m,1H), 2.18 (dd, J=13.0, 11.7 Hz, 1H), 2.04 (dd, J=13.0, 7.2 Hz, 1H),1.88-1.79 (m, 1H), 1.79-1.70 (m, 1H). Peak 2: 1.8 g (97.2% opticalpurity at 222 nm). Ret. Time=7.71 min on analytical chiral SFC. ¹H NMR(500 MHz, METHANOL-d₄) δ 7.45-7.38 (m, 2H), 7.26-7.20 (m, 2H), 3.78 (s,3H), 3.28-3.20 (m, 1H), 2.66-2.57 (m, 1H), 2.25 (ddd, J=12.8, 11.0, 7.2Hz, 1H), 2.10 (dt, J=12.2, 6.8 Hz, 1H), 2.03-1.93 (m, 1H), 1.84 (ddd,J=13.0, 7.8, 2.2 Hz, 1H), 1.65 (dd, J=13.3, 11.1 Hz, 1H).

Intermediates 3-I and 3-II (1R,3S)-methyl1-amino-3-(4-iodophenyl)cyclopentanecarboxylate hydrochloride (I-3I) and(1R,3R)-methyl 1-amino-3-(4-iodophenyl)cyclopentanecarboxylateHydrochloride

Intermediate 3A: 3-(4-iodophenyl)cyclopentanone

To a solution of cyclopent-2-enone (3.39 g, 0.0407 mol), sodium acetate(6.659 g, 0.0813 mol) and (4-iodophenyl)boronic acid (10 g, 0.0407 mol)in acetic acid (325 mL) was added palladium (II) acetate (0.9728 g,0.00406 mol) and antimony (III) chloride (0.9279 g, 0.00406 mol) under anitrogen atmosphere. After being stirred for 2 hours at 25° C., theblack precipitation was filtered off and the filtrate was diluted withbrine and then extracted twice with dichloromethane. The organicextraction was stirred with saturated sodium bicarbonate for 30 minutes,then washed with brine and dried over sodium sulfate. Removal of thesolvent resulted in a yellow oil. Further purification (flash column,chloroform eluent) gave a 6.5 g of 3-(4-iodophenyl)cyclopentanone as awhite solid.

Step B: Methyl 1-amino-3-(4-iodophenyl)cyclopentanecarboxylate

To a solution of 3-(4-iodophenyl)cyclopentanone (10 g, 0.03496 mol) inmethanolic ammonia (7 M, 105 mL) was added sodium cyanide (3.42 g,0.06993 mmol) and ammonium chloride (3.74 g, 0.06993 mmol). The mixturewas allowed to stir at room temperature for 72 hours. Aqueous sodiumbicarbonate was added and the reaction mixture was extracted with ethylacetate. The organic layer was evaporated over sodium sulfate in thepresence of vacuum distillation and the obtained crude compound was usedas such for the subsequent reaction. The crude mixture of1-amino-3-(4-iodophenyl) cyclopentanecarbonitrile was dissolved inconcentrated hydrochloric acid and refluxed at 70° C. overnight. Thereaction mixture was distilled and then co-distilled with water. Acetonewas added to the reaction, which was stirred for 30 minutes and theresulting solids (7 g) were filtered off. The solids were used directlyin the subsequent step. The solids were dissolved in methanol (140 mL)and thionyl chloride (19.9 g, 0.169184 mol) was added under nitrogen, inthe presence of ice water bath cooling. The reaction mixture was thenallowed to stir at 70° C. overnight. The methanol was removed bydistillation and aqueous sodium bicarbonate was added. The solution wasextracted with ethyl acetate. The solution was dried over sodium sulfateand concentrated to provide the product (4.5 g) as a brown oil.

Step C: Intermediates 3I and 3II

Methyl 1-amino-3-(4-iodophenyl)cyclopentanecarboxylate) (approximately 5g) was purified by Chiral SFC under the conditions described below. Thefour isomers were isolated and named “Pk1”, “Pk2”, and “Pk3” and “Pk4”in the elution order. The diastereoisomeric purity of each isolateisomer was determined on the SFC/UV/area % at 220 nm and summarizedbelow. The methanol was evaporated to give the four individual isomersas reddish brown oils. Based on the proton NMR data, Peaks 1 and 4 wereenantiomeric and Peaks 2 and 3 were enantiomeric. Absolute configurationwas established through correlation to Intermediate 1A and Intermediate1B after conversion to common products. Instrument: Berger SFC MGIII.Preparative Conditions: Column: ChiralPak AD-H 25×5 cm, 5 μm; ColumnTemp. 35° C.; Flow rate: 135 mL/min; Mobile Phase: CO₂/(MeOH+0.5%DEA)=65/35; Injection Vol. 0.7 mL; Detector Wavelength 220 nm. SampleConc. (mg/mL) 30 mg/mL.

Diastereoisomeric purity (Area %) of each isomer Pk1 Pk2 Pk3 Pk4Diastereoisomeric 97.5% 96.3% 95.6% 95.6% purity (Area %) Weight  1.178g  1.372 g  1.312 g  1.216 g

Intermediate 3I (Pk1): HPLC retention time=10.62 min (condition I);LC/MS M⁺¹=346.0. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.71-7.55 (m, J=8.1 Hz,2H), 7.16-7.02 (m, J=8.1 Hz, 2H), 3.76 (s, 3H), 3.28-3.10 (m, 1H), 2.61(dd, J=13.2, 7.9 Hz, 1H), 2.31-2.19 (m, 1H), 2.15-2.05 (m, 1H),2.03-1.91 (m, 1H), 1.88-1.79 (m, 1H), 1.64 (dd, J=13.1, 11.3 Hz, 1H).

Intermediate 3II (Pk3): HPLC retention time=10.64 min (condition I);LC/MS M⁺¹=346.0. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.71-7.56 (m, J=8.4 Hz,2H), 7.18-7.01 (m, J=8.1 Hz, 2H), 3.75 (s, 3H), 3.49-3.36 (m, 1H), 2.40(ddd, J=12.7, 8.8, 3.4 Hz, 1H), 2.29-2.13 (m, 2H), 2.10-1.99 (m, 1H),1.90-1.69 (m, 2H).

Intermediate 4(5R,7S)-7-(4-bromophenyl)-3-oxa-1-azaspiro[4.4]nonan-2-one

Intermediate 4A: ((1R,3S)-1-amino-3-(4-bromophenyl)cyclopentyl)methanol

To a mixture of (1R,3S)-methyl 1-amino-3-(4-bromophenyl)cyclopentanecarboxylate, HCl (15 g, 44.8 mmol) in MeOH (100 mL) at 0° C.was added sodium borohydride (4 g, 106 mmol) portionwise. The reactionmixture was warmed to room temperature and sodium borohydride was addedportionwise until the reaction was determined to be complete by HPLCanalysis. Water was added to quench the reaction. The reaction mixturewas diluted with ethyl acetate and washed with saturated NaCl. Theaqueous layer was back extracted several times. The combined organiclayers were dried with MgSO₄, filtered and concentrated. The product (11g) was recovered after concentration. HPLC retention time=0.65 min(condition G); LC/MS M⁺¹=272: ¹H NMR (400 MHz, DMSO-d₆) δ 7.51-7.40 (m,2H), 7.27 (d, J=8.4 Hz, 2H), 3.32-3.20 (m, 2H), 3.09-2.92 (m, 1H), 2.11(dd, J=12.9, 8.7 Hz, 1H), 1.98-1.87 (m, 1H), 1.80 (qd, J=11.1, 7.9 Hz,1H), 1.69-1.58 (m, 1H), 1.48 (ddd, J=12.4, 7.9, 2.2 Hz, 1H), 1.32 (dd,J=12.8, 10.1 Hz, 1H).

Intermediate 4

To a mixture of ((1R,3S)-1-amino-3-(4-bromophenyl)cyclopentyl)methanol(11 g, 40.7 mmol) and pyridine (3.29 mL, 40.7 mmol) in dioxane (300 mL)was added 1,1′-carbonyldiimidazole (19.81 g, 122 mmol). The reactionmixture was stirred for 4 hours. The reaction mixture was diluted withethyl acetate and washed with 1M HCl, brine and saturated NaHCO₃. Themixture was back extracted several times. The organic layer was driedwith MgSO₄, filtered and concentrated to afford 10.5 g of desiredproduct as an off-white solid. HPLC retention time=0.87 min (conditionG). LC/MS M⁺¹=297.9; ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.45 (d, J=8.6 Hz,2H), 7.12 (d, J=8.4 Hz, 2H), 6.42 (br. s., 1H), 4.41-4.21 (m, 2H),3.17-2.91 (m, 1H), 2.34 (dd, J=13.3, 7.4 Hz, 1H), 2.23-2.11 (m, 2H),2.01-1.90 (m, 2H), 1.88-1.74 (m, 1H).

Intermediate 5(5R,7R)-7-(4-bromophenyl)-3-oxa-1-azaspiro[4.4]nonan-2-one

Intermediate 5A: ((1R,3R)-1-amino-3-(4-bromophenyl)cyclopentyl)methanol

(1R,3R)-methyl 1-amino-3-(4-bromophenyl)cyclopentanecarboxylate (3.88 g,13.01 mmol) was dissolved in MeOH (65.1 ml) and sodium borohydride(1.477 g, 39.0 mmol) was added portion wise. Additional sodiumborohydride was added (0.5 equiv every 1 h) portion wise until thereaction was determined to be complete by HPLC analysis. The reactionwas found to be complete after 2 hours. The reaction mixture wasquenched with water and diluted with ethyl acetate. The aqueous layerwas back extracted three times with EtOAc. The organic layers werecombined, washed with saturated NaCl, dried over MgSO₄, filtered andconcentrated to afford((1R,3R)-1-amino-3-(4-bromophenyl)cyclopentyl)methanol (3.19 g, 11.81mmol). HPLC ret time=0.68 min (cond); LC/MS M⁺¹=272: ¹H NMR (400 MHz,CDCl₃) δ 7.42 (d, J=8.4 Hz, 2H), 7.13 (d, J=8.4 Hz, 2H), 3.49 (s, 2H),3.32-3.41 (m, 1H), 2.19-2.25 (m, 1H), 1.98-2.07 (m, 1H), 1.90-1.95 (m,1H), 1.66-1.74 (m, 2H), 1.52-1.60 (m, 1H).

Intermediate 5

((1R,3R)-1-amino-3-(4-bromophenyl)cyclopentyl)methanol (3.19 g, 11.81mmol) was dissolved in THF (59.0 ml). Pyridine (0.955 ml, 11.81 mmol)and 1,1′-carbonyldiimidazole (5.74 g, 35.4 mmol) were added portionwise. The reaction mixture was stirred for 4 h and was followed by LCMS.After completion, the mixture was diluted with EtOAc and washed with 1MHCl. The aqueous layer was back extracted twice with EtOAc. The organiclayers were combined, washed with saturated NaCl, dried over MgSO₄,filtered and concentrated to afford(5R,7R)-7-(4-bromophenyl)-3-oxa-1-azaspiro[4.4]nonan-2-one (2.5 g, 8.44mmol) after flash chromatography (24 g silica gel column; eluent: hexane2 CV followed by a gradient to 100% EtOAc over 15 CV) HPLC ret time=0.91min (cond); LC/MS M⁺¹=298. ¹H NMR (400 MHz, CDCl₃) δ 7.46 (d, J=8.5 Hz,2H), 7.09 (d, J=8.5 Hz, 2H), 5.72-5.81 (m, 1H), 4.35 (dd, J=13 Hz, 8 Hz,2H), 3.19-3.24 (m, 1H), 2.38-2.44 (m, 1H), 2.15-2.26 (m, 1H), 2.11-2.14(m, 1H), 1.99-2.05 (m, 1H), 1.79-1.85 (m, 1H), 1.65-1.72 (m, 1H).

Intermediate 6 Methyl1-((diphenylmethylene)amino)cyclopent-3-enecarboxylate

To a mixture of (N,N-diphenylmethylgene)glycine ethyl ester (4 g, 14.96mmol) in THF (3 mL) at 0° C. was added lithium bis(trimethylsilyl)amide(16.46 mL, 16.46 mmol) dropwise over 30 minutes. After stirring for 30minutes, the resulting solution was then added dropwise tocis-1,4-dichloro-2-butene (1.823 mL, 16.46 mmol) in THF (1 mL). After 1hour, lithium bis(trimethylsilyl)amide (14.96 mL, 14.96 mmol) was addedat 0° C. The mixture was stirred at room temperature for 8 hours beforebeing quenched by saturated aqueous NH₄Cl solution (30 mL) and water (10mL). The mixture was extracted with ethyl acetate (3×20 mL). Thecombined ethyl acetate extracts were dried (Na₂SO₄) and concentratedunder reduced pressure. The crude material was filtered through a shortplug of silica and purified by silica gel chromatography. HPLC retentiontime=5.06 min (condition H); LC/MS M⁺¹=320. ¹H NMR (400 MHz,METHANOL-d₄) δ 7.23-7.18 (m, 2H), 7.17-7.10 (m, 2H), 3.72-3.56 (m, 2H),3.24-3.08 (m, 1H), 2.71 (s, 3H), 2.60 (t, J=7.6 Hz, 2H), 2.44 (ddd,J=13.4, 7.1, 1.3 Hz, 1H), 2.18 (t, J=7.6 Hz, 3H), 2.03-1.89 (m, 3H),1.75 (t, J=12.8 Hz, 1H), 1.69-1.56 (m, 4H), 1.45-1.27 (m, 2H).

Intermediate 7(5R,7S)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

Intermediate 7A: tert-butyl2-(4-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl) phenyl)acetate

To a mixture of(5R,7S)-7-(4-bromophenyl)-3-oxa-1-azaspiro[4.4]nonan-2-one (1 g, 3.38mmol) in dioxane (10 mL) at room temperature was added lithiumbis(trimethylsilyl)amide (3.71 mL, 3.71 mmol). The mixture was stirredfor 30 minutes, then1,2,3,4,5-pentaphenyl-1′-(di-t-butylphosphino)ferrocene (0.121 g, 0.169mmol), Pd₂(dba)₃ (0.155 g, 0.169 mmol) and(2-(tert-butoxy)-2-oxoethyl)zinc(II) chloride (8.10 mL, 4.05 mmol) wereadded. The reaction mixture was heated at 80° C. for 2 hours, thencooled to room temperature, diluted with ethyl acetate and washed with1M HCl. The organic layer was dried with MgSO₄, filtered andconcentrated. The crude material was purified on a silica gel cartridge(40 g) using an EtOAc/hexane gradient (0-100% EtOAc over 20 minutes) toafford 950 mg of tert-butyl2-(4-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)phenyl)acetate. HPLCretention time=0.93 min (condition G); LC/MS M⁺¹=332.

Intermediate 7B:2-(4-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)phenyl)acetic Acid

To a mixture of tert-butyl2-(4-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl) phenyl)acetate (1g, 3.02 mmol) in DCM (20 mL) was added TFA (10 mL). After 2 h, thesolution was concentrated in vacuo and used as such for the subsequentstep without further purification. HPLC retention time=0.65 min(condition G); LC/MS M⁺¹=276.

Intermediate 7

To a mixture of2-(4-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)phenyl) acetic acid(800 mg, 2.91 mmol) in DCM (20 mL) was added oxalyl chloride (1 ml,11.42 mmol) and a few drops of DMF. After one hour, the reaction mixturewas concentrated in vacuo. The residue was re-dissolved in DCM (20 mL)in a glass pressure vessel. Granular aluminum chloride (1550 mg, 11.62mmol) was added and the reaction mixture was cooled to −78° C. Ethylenewas bubbled through the solution for 5 minutes and then the reactionvessel was sealed. The reaction mixture was allowed to slowly warm toroom temperature and stirred for 4 hours. The mixture was poured ontoice, diluted with dichloromethane and washed with 1M HCl. The organiclayer was dried with MgSO₄, filtered and concentrated. The crudematerial was purified on a silica gel cartridge (80 g) using a MeOH/DCMgradient (0-10% MeOH over 13CV). The product containing fractions werecollected and dried in vacuo to afford 770 mg of(5R,7S)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.HPLC retention time=0.74 min (condition G); LC/MS M⁺¹=286: ¹H NMR (400MHz, CHLOROFORM-d) δ 7.20-7.00 (m, 3H), 5.49 (br. s., 1H), 4.45-4.25 (m,2H), 3.59 (s, 2H), 3.08 (t, J=6.8 Hz, 3H), 2.58 (t, J=6.7 Hz, 2H), 2.38(dd, J=13.2, 7.3 Hz, 1H), 2.27-2.11 (m, 2H), 2.05-1.92 (m, 2H),1.92-1.74 (m, 1H).

Intermediate 86-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-3,4-dihydronaphthalen-2-yltrifluoromethanesulfonate

To a mixture of(5R,7S)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(340 mg, 1.192 mmol) and DMPU (0.431 mL, 3.57 mmol) in THF (10 mL) at−78° C. was added LDA (1.456 mL, 2.62 mmol). The reaction mixture wasstirred for 30 minutes then1,1,1-trifluoro-N-phenyl-N-(trifluoromethyl)sulfonyl methanesulfonamide(639 mg, 1.787 mmol) in THF (10 mL) was added. The reaction mixture waswarmed to 0° C. After 1 hour, the reaction was quenched with water. Thereaction mixture was diluted with ethyl acetate and washed withsaturated aqueous NaCl. The organic layer was dried with MgSO₄, filteredand concentrated. The crude material was purified on a silica gelcartridge (40 g) using an EtOAc/hexane gradient (0-100% EtOAc over 20minutes) to afford 400 mg of6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-3,4-dihydronaphthalen-2-yltrifluoromethanesulfonate. HPLC retention time=1.01 min (condition G);LC/MS M⁺¹=418. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.17-6.95 (m, 3H), 6.74(s, 1H), 6.48 (s, 1H), 4.48-4.20 (m, 2H), 3.17-2.95 (m, 3H), 2.81-2.60(m, 2H), 2.33 (dd, J=13.3, 7.2 Hz, 1H), 2.24-2.08 (m, 2H), 2.05-1.74 (m,3H).

Intermediate 96-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-3,4-dihydronaphthalen-2-yltrifluoromethanesulfonate

To a mixture of(5R,7R)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(250 mg, 0.876 mmol) and DMPU (317 μl, 2.63 mmol) in THE (10 mL) at −78°C. was added LDA (1071 μl, 1.928 mmol). The reaction mixture was stirredfor 30 minutes then 1,1,1-trifluoro-N-phenyl-N-(trifluoromethyl)sulfonylmethanesulfonamide (470 mg, 1.314 mmol) in THF (4381 μl) was added. Thereaction mixture was warmed to 0° C. and stirred for 1 hour. LCMS showedconversion to be complete. The reaction was quenched with water. Thereaction mixture was diluted with ethyl acetate and washed withsaturated NaCl. The organic layer was dried Na₂SO₄, filtered andconcentrated under reduced pressure. The crude material was purified ona silica gel cartridge (80 g) using an EtOAc/Hex gradient (0-100% EtOAcover 12 CV). Product containing fractions were combined, concentrated,and dried in vacuo to afford6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-3,4-dihydronaphthalen-2-yltrifluoromethanesulfonate (200 mg, 0.479 mmol). HPLC retention time=1.12min (condition G) LC/MS M⁺¹=418.3.

Intermediate 10(5R,7R)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

Intermediate 10A: tert-butyl2-(4-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl) phenyl)acetate

To a solution of(5R,7R)-7-(4-bromophenyl)-3-oxa-1-azaspiro[4.4]nonan-2-one (Int. 4, 2.1g, 7.09 mmol) in THF (25.3 ml) at room temperature was added LiHMDS(7.80 ml, 7.80 mmol). The solution was stirred for 15 min. Next,Pd₂(dba)₃ (0.195 g, 0.213 mmol),1,2,3,4,5-pentaphenyl-1′-(di-t-butylphosphino)ferrocene (0.151 g, 0.213mmol), and (2-(tert-butoxy)-2-oxoethyl)zinc(II) bromide, tetrahydrofuran(7.07 g, 21.27 mmol) were sequentially added. The slurry was stirred at24° C. for 2 h. LCMS analysis showed complete consumption of thestarting material. The reaction mixture was diluted with ethyl acetateand washed with 1M HCl. The organic layer was dried over MgSO₄, filteredand concentrated. The crude material was purified on a silica gelcartridge (40 g) using hexane: acetone 100:0 to 0:100 over 25 CV.Tert-butyl2-(4-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)phenyl)acetate (2.35g, 7.09 mmol) was isolated. HPLC retention time=0.95 min (condition I):LC/MS M⁺¹=332: ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.27-7.21 (m, 2H),7.21-7.15 (m, 2H), 5.11 (br. s., 1H), 4.40-4.26 (m, 2H), 3.53 (s, 2H),3.22-3.01 (m, 1H), 2.36 (dd, J=13.2, 7.3 Hz, 1H), 2.25-2.10 (m, 2H),2.04-1.92 (m, 2H), 1.91-1.76 (m, 1H), 1.47 (s, 9H).

Intermediate 10

The brown liquid tert-butyl2-(4-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)phenyl)acetate (2.35g, 7.09 mmol) was dissolved in DCM (60 mL) followed by the addition oftrifluoroacetic acid (20 mL, 260 mmol). The reaction mixture was stirredat room temperature for 1 h at which time the solvent was removed underreduced pressure. The resulting material was diluted in DCM (60 mL),purified by acid/base extraction and placed under vacuum for 1 h. Theresulting brown gum2-(4-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)phenyl)acetic acid(1.952 g, 7.09 mmol) was dissolved in DCM (60 mL) followed by theaddition of oxalyl chloride (1.862 mL, 21.27 mmol), and DMF (0.027 mL,0.355 mmol). The resulting solution was stirred until the evolution ofgas ceased (about 30 min) at room temperature. LCMS of an aliquotquenched with MeOH showed complete consumption of the acid (RT=0.65 min,Condition I) and appearance of the presumed methyl ester due to methanolquench (RT=0.77 min, condition I) as the only product. The solvent wasremoved under reduced pressure and the product was placed under vacuum.The brown gum was transfer to a sealed tube with DCM (60 mL) (does notcompletely dissolve, a brown suspension was obtained). The reactionmixture was cooled to −78° C. followed by the addition of granularaluminum chloride (2.84 g, 21.27 mmol). Ethylene was bubbled through thesolution for 7 min and the tube was sealed. A precipitate formed and thereaction mixture was stirred at −78° C. for 15 min and then allowed toreach room temperature. The reaction mixture was stirred for 2 h at roomtemperature and then depressurized. LCMS analysis showed disappearanceof starting material and appearance of the tetralone product. Thereaction mixture was poured over ice, diluted with DCM and stirred untilthe ice melted. The organic layer was washed with brine, dried andconcentrated under reduced pressure. Purification on silica gel afforded(5R,7R)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(1.05 g, 3.68 mmol). HPLC retention time=0.74 min (condition I); LC/MSM⁺¹=286; ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.23-7.11 (m, 3H), 5.68 (br.s., 1H), 4.45-4.30 (m, 2H), 3.59 (s, 2H), 3.31-3.18 (m, 1H), 3.08 (t,J=6.8 Hz, 2H), 2.58 (t, J=6.7 Hz, 2H), 2.42-2.39 (m, 1H), 2.32-2.15 (m,2H), 2.09-1.99 (m, 1H), 1.91-1.83 (m, 1H), 1.82-1.72 (m, 1H).

Intermediate 12 6-bromo-2-hexyl-3,4-dihydroisoquinolin-1(2H)-one

To a stirred cloudy solution of 6-bromo-3,4-dihydro-2H-isoquinolin-1-one(0.6 g, 2.65 mmol) and 1-iodohexane (0.783 mL, 5.31 mmol) in anhydroustetrahydrofuran (30 mL) was added 60% mineral oil dispersion of sodiumhydride (0.212 g, 5.31 mmol) portionwise over 20 min. The reactionmixture was stirred at room temperature under nitrogen for 1 h and 65°C. for 3 h. Additional sodium hydride (0.25 g) and 1-iodohexane (1 mL)were added. The mixture was stirred at 65° C. for 1 h. After anhydrousDMF (3 mL) was added at room temperature, the mixture was stirred atroom temperature for 2.5 days. The reaction was quenched with saturatedaqueous ammonium chloride solution (6 mL) and water (3 mL). Hexanes (20mL) were added. The organic solution was separated and washed with water(10 mL). The combined aqueous solutions were extracted with ethylacetate (3×5 mL). The combined organic solutions were dried overanhydrous sodium sulfate and concentrated. Flash chromatographypurification (24 g silica gel column, gradient elution from 0 to 50%ethyl acetate in hexanes) afforded6-bromo-2-hexyl-3,4-dihydroisoquinolin-1(2H)-one (667 mg, 2.150 mmol) asa yellowish solid. LC/MS M⁺¹=310, 312. ¹H NMR (400 MHz, CHLOROFORM-d) δ7.94 (d, J=8.1 Hz, 1H), 7.46 (dd, J=8.3, 1.9 Hz, 1H), 7.36-7.30 (m, 1H),3.60-3.46 (m, 4H), 2.96 (t, J=6.6 Hz, 2H), 1.68-1.57 (m, 2H), 1.32 (br.s., 6H), 0.93-0.83 (m, 3H).

Intermediate 13 6-bromo-2-(pentyloxy)-1,2,3,4-tetrahydronaphthalene

Step A: 6-bromo-1,2,3,4-tetrahydronaphthalen-2-ol

To a stirred solution of 6-bromo-3,4-dihydronaphthalen-2(1H)-one (2.00g, 8.89 mmol) in ethanol (15 mL) and dichloromethane (5 mL) was addedsodium borohydride (0.336 g, 8.89 mmol) portionwise at room temperatureunder nitrogen. The mixture was stirred at room temperature overnight.The reaction was quenched with acetone (2 mL). After being stirred atroom temperature for 1 h, the mixture was concentrated. The residue waspartitioned between saturated aqueous ammonium chloride solution (5 mL),water (3 mL), and ethyl acetate (10 mL). The aqueous layer was separatedand extracted with ethyl acetate (3×3 mL). The combined ethyl acetatesolutions were dried (anhydrous sodium sulfate) and concentrated underreduced pressure. Flash chromatography purification (40 g silica gelcolumn, gradient elution from 5 to 100% ethyl acetate in hexanes)afforded 6-bromo-1,2,3,4-tetrahydronaphthalen-2-ol (1.55 g, 6.83 mmol)as a liquid. LC/MS [M−H₂O]⁺¹=209, 211. Chiral SFC separation (AD-H (5×25cm), 15% MeOH in CO₂, 300 ml/min, 220 nm, 35° C.) gave PK1 (560 mg) andPK2 (580 mg) as yellow liquids. Both isomers were converted to theiramyl ethers as shown below.

Step B: Intermediate 13

To a stirred solution of 6-bromo-1,2,3,4-tetrahydronaphthalen-2-ol (0.58g, 2.55 mmol) (PK2) in anhydrous tetrahydrofuran (20 mL) was added 60%mineral oil dispersion of sodium hydride (0.511 g, 12.77 mmol)portionwise. The mixture was stirred at room temperature for 15 minbefore n-amyl iodide (1.340 mL, 10.22 mmol) was added. The mixture wasstirred at room temperature under nitrogen for two days. More 60%mineral oil dispersion of sodium hydride (0.511 g, 12.77 mmol), n-amyliodide (1.340 mL, 10.22 mmol), and anhydrous tetrahydrofuran (20 mL)were added and the mixture was stirred at room temperature over twodays. Saturated aqueous ammonium chloride solution (9 mL) was addedslowly. The mixture was concentrated. The aqueous residue was extractedwith ethyl acetate (4×5 mL). The combined ethyl acetate extracts weredried (anhydrous sodium sulfate) and concentrated under reduced pressureto give a liquid. Flash chromatography purification (120 g silica gelcolumn, gradient elution from 0 to 5% ethyl acetate in hexanes) afforded6-bromo-2-(pentyloxy)-1,2,3,4-tetrahydronaphthalene (0.61 g, 2.052 mmol)as a yellow liquid. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.24-7.18 (m, 2H),6.93 (d, J=7.9 Hz, 1H), 3.75-3.66 (m, 1H), 3.57-3.45 (m, 2H), 3.04-2.85(m, 2H), 2.79-2.66 (m, 2H), 2.08-1.98 (m, 1H), 1.85-1.74 (m, 1H),1.64-1.55 (m, 2H), 1.39-1.25 (m, 4H), 0.95-0.85 (m, 3H).

Intermediate 14 6-bromo-2-hexylchroman

Step A: 6-bromo-2-hexylchroman-4-one

To a stirred solution of 5′-bromo-2′-hydroxyacetophenone (3.2 g, 14.88mmol) and n-heptaldehyde (2.199 mL, 16.37 mmol) in methanol (50 mL) wasadded pyrrolidine (2.484 mL, 29.8 mmol) at room temperature undernitrogen. The mixture was stirred at 70° C. for 2 h and at roomtemperature overnight. The solvent was evaporated. Flash chromatographypurification (330 g silica gel column; gradient elution from 0 to 10%ethyl acetate in hexanes) afforded 6-bromo-2-hexylchroman-4-one (3.58 g,11.50 mmol) as a liquid.

Step B: Intermediate 14

To a stirred solution of 6-bromo-2-hexylchroman-4-one (1.8 g, 5.78 mmol)in ethanol (10 mL) was added sodium borohydride (0.109 g, 2.89 mmol).The resulting mixture was stirred at room temperature under nitrogen for2 hr before being concentrated. The residue was mixed with saturatedaqueous ammonium chloride solution (5 mL), water (3 mL), and ethylacetate (6 mL). The aqueous layer was separated and extracted with ethylacetate (3×3 mL). The combined ethyl acetate extracts were dried overNa₂SO₄ and concentrated under reduced pressure to give a liquid.

The liquid was mixed with triethylsilane (4.62 mL, 28.9 mmol).Trifluoroacetic acid (2.228 mL, 28.9 mmol) was added dropwise at roomtemperature under nitrogen. The mixture was stirred vigorously at roomtemperature for 2 hr before water (10 mL) was added. The aqueous layerwas separated and extracted with a mixture of ethyl acetate and hexanes(1:1; 3×3 mL). The combined organic solutions were washed water and thenwith saturated aqueous sodium bicarbonate solution until it was basic,dried (Na₂SO₄), and concentrated under reduced pressure. Flashchromatography purification using ISCO (40 g silica gel column, 0% to30% ethyl acetate in hexanes over 15 min) afforded6-bromo-2-hexylchroman (1.4 g, 4.71 mmol) as a yellow liquid. ¹H NMR(400 MHz, CHLOROFORM-d) δ 7.18-7.11 (m, 2H), 6.67 (d, J=9.2 Hz, 1H),3.95 (dddd, J=9.8, 7.3, 5.1, 2.2 Hz, 1H), 2.85-2.66 (m, 2H), 1.97 (dddd,J=13.6, 5.9, 3.5, 2.3 Hz, 1H), 1.79-1.21 (m, 11H), 0.93-0.85 (m, 3H).Chiral SFC separation (Chiralcel OJ-H 3×250 cm, 5 um; CO₂/IPA=95/5; 180mL/min; 220 nm) of the liquid afforded Isomer 1 (0.37 g) and Isomer 2(0.4 g) as yellow liquids.

Intermediate 15 methyl6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate

Intermediate 15A: Methyl4-oxo-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate

To a mixture of potassium carbonate (523 mg, 3.78 mmol),(5R,7S)-7-(4-bromophenyl)-3-oxa-1-azaspiro[4.4]nonan-2-one (800 mg, 2.70mmol), itaconic acid (457 mg, 3.51 mmol), and acetonitrile (8 mL) wasadded water (2.4 mL). The mixture was stirred till the evolution ofcarbon dioxide stopped and then bubbled with nitrogen for 3 min. Afterpalladium(II) acetate (30.3 mg, 0.135 mmol) and tri-o-tolylphosphine (82mg, 0.270 mmol) were added, the mixture was bubbled with nitrogen for anadditional 3 min. The mixture was stirred at 80° C. for 20 h and thenconcentrated. The residue was mixed with water (40 mL), basified withpotassium carbonate and filtered. The filtrate was washed with diethylether (2×15) then acidified to pH approximately 2 with 6N aqueoushydrochloric acid. The solid was separated and the aqueous solution wasextracted with a mixture of THF/EtOAc (3:1) (4×10 mL). The solid and theextracts were combined and concentrated. LC/MS [M−H₂O]⁺¹=328.

The residue was mixed with THF (5 mL), ethyl acetate (5 mL), methanol(20 mL), and 10% Pd/C (400 mg, 0.376 mmol) and hydrogenated under ahydrogen balloon overnight. The catalyst was filtered off through amembrane filter and washed with methanol. The filtrate was concentratedand lyophilized to give a solid. LC/MS [M−H₂O]⁺¹=330.

The solid was mixed with 98% sulfuric acid (15 mL, 281 mmol). The clearsolution was stirred at room temperature for 4 h. Methanol (8 mL, 198mmol) was added slowly with water-bath cooling. The mixture was stirredat room temperature for 1 h before being poured onto ice (150 g). Themixture was extracted with ethyl acetate (4×40 mL). The combined ethylacetate extracts were washed with saturated aqueous sodium bicarbonatesolution (20 mL), dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. Flash chromatography purification (24 g silicagel column, gradient elution from 10 to 100% ethyl acetate in hexanes)afforded methyl4-oxo-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(440 mg, 1.281 mmol). LC/MS M⁺¹=344.

Intermediate 15

A mixture of methyl4-oxo-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(440 mg, 1.281 mmol), MeOH (15 mL), acetic acid (1.5 mL), and 10% Pd/C(200 mg, 0.188 mmol) was hydrogenated under a hydrogen balloon over aweekend. The mixture was filtered through a membrane filter. Thefiltrate was concentrated to give methyl6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(337 mg, 1.023 mmol) as a white solid. LC/MS M⁺¹=330.

Intermediates 16-I and 16-II(5R,7S)-7-((S)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(I-16-I) and(5R,7S)-7-((R)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(I-16-II)

A mixture of methyl6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(337 mg, 1.023 mmol), anhydrous tetrahydrofuran (3 mL), and 2N THFsolution of lithium borohydride (2.56 mL, 5.12 mmol) was stirred at 70°C. for 4 h. Saturated aqueous ammonium chloride solution was addedslowly at 0° C. to quench the reaction. Water and ethyl acetate wereadded. The aqueous solution was extracted with ethyl acetate. Thecombined ethyl acetate solutions were dried over anhydrous sodiumsulfate and concentrated under reduced pressure to give(5R,7S)-7-(6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(300 mg, 0.995 mmol). LC/MS M⁺¹=302.

Chiral SFC separation (AD-H (0.46×25 cm), 45% MeOH in CO₂, 3 ml/min, 220nm, 35° C.) gave enantiomers 1 and 2 as white solids. Isomer 1:Intermediate 16-I(5R,7S)-7-((S)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.HPLC retention time=2.87 min (condition C); LC/MS M⁺¹=330. ¹H NMR (400MHz, CHLOROFORM-d) δ 7.06 (d, J=7.9 Hz, 1H), 6.96 (d, J=7.8 Hz, 1H),6.92 (s, 1H), 5.22 (br. s., 1H), 4.38-4.18 (m, 2H), 3.73 (s, 3H),3.07-2.91 (m, 3H), 2.89-2.79 (m, 2H), 2.78-2.68 (m, 1H), 2.31 (dd,J=13.3, 7.3 Hz, 1H), 2.25-2.16 (m, 1H), 2.16-2.07 (m, 2H), 2.01-1.76 (m,4H). Isomer 2: Intermediate 16-II(5R,7S)-7-((R)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one;HPLC retention time=2.88 min (condition C); LC/MS M⁺¹=330; ¹H NMR (400MHz, CHLOROFORM-d) δ 7.06 (d, J=7.9 Hz, 1H), 6.96 (d, J=7.9 Hz, 1H),6.92 (s, 1H), 5.05 (br. s., 1H), 4.35-4.24 (m, 2H), 3.73 (s, 3H),3.08-2.92 (m, 3H), 2.90-2.79 (m, 2H), 2.78-2.68 (m, 1H), 2.32 (dd,J=13.3, 7.2 Hz, 1H), 2.25-2.06 (m, 3H), 2.01-1.74 (m, 4H).

Intermediate 17-I and 17-II((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (I-17-I) and((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (I-17-II)

Int-17-II:(5R,7S)-7-((R)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(enantiomer 2; 690 mg, 2.289 mmol) was dissolved in dry pyridine (5 mL)and p-toluenesulfonyl chloride (1309 mg, 6.87 mmol) was added in oneportion. The resulting mixture was reacted at room temperature for 4 h.The solvent was removed in vacuo. The residue was dissolved in methylenechloride and methanol. Flash chromatography purification (40 g silicagel column, gradient elution from 20 to 100% ethyl acetate in hexanes)afforded((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (860 mg, 1.888 mmol) as a white solid. LC/MSM⁺¹=456.Int-17-I was prepared according to the general procedure as Intermediate17-II using(5R,7S)-7-((S)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-oneto afford((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate.

Examples 1 to 4(1-amino-3-((R)-2-((pentyloxy)methyl)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)cyclopentyl)methanol

Preparation 1A: (S)-1-(4-bromo-2-(oxiran-2-ylmethoxy)phenyl)ethanone

To a stirred solution of 4-bromo-2-hydroxyacetophenone (2.54 g, 11.81mmol) in anhydrous DMF (10 mL) was added sodium hydride (60% dispersionin mineral oil, 0.520 g, 12.99 mmol). The mixture was stirred at roomtemperature for 40 min before a solution of(2S)-glycidyl-3-nitrobenzenesulfonate (3.37 g, 12.99 mmol) in anhydrousDMF (5 mL) was added dropwise at room temperature under nitrogen. Themixture was stirred at 70° C. for 3.5 h. The reaction mixture wasconcentrated to remove DMF and the residue was quenched with 10% aqueouscitric acid to pH approximately 3. The mixture was diluted with water (5mL) and extracted with ethyl acetate (4×10 mL). The combined extractswere dried over anhydrous sodium sulfate and concentrated. Flashchromatography purification (80 g silica gel column, gradient elutionfrom 0 to 40% ethyl acetate in hexanes) afforded(S)-1-(4-bromo-2-(oxiran-2-ylmethoxy)phenyl)ethanone (2.68 g, 9.89 mmol)as a white solid. LC/MS M⁺²³=293, 295. ¹H NMR (400 MHz, CHLOROFORM-d) δ7.64 (d, J=8.4 Hz, 1H), 7.19 (dd, J=8.3, 1.7 Hz, 1H), 7.12 (d, J=1.8 Hz,1H), 4.40 (dd, J=10.9, 2.8 Hz, 1H), 4.00 (dd, J=11.0, 6.2 Hz, 1H),3.45-3.38 (m, 1H), 3.00-2.93 (m, 1H), 2.78 (dd, J=4.8, 2.6 Hz, 1H), 2.65(s, 3H).

Preparation 1B: (S)-4-bromo-2-(oxiran-2-ylmethoxy)phenylacetate

To a stirred solution of(S)-1-(4-bromo-2-(oxiran-2-ylmethoxy)phenyl)ethanone (0.76 g, 2.80 mmol)in methylene chloride (30 mL) were added sodium bicarbonate (1.6 g,19.05 mmol) and m-CPBA (1.257 g, 5.61 mmol). The mixture was stirred at40° C. for 5 h and room temperature overnight. The solid was filteredoff and the filtrate was concentrated. The residue was dissolved inethyl acetate, washed with a mixture of sodium bicarbonate andthiosulfate aqueous solutions, dried over anhydrous sodium sulfate, andconcentrated. Flash chromatography purification (24 g silica gel column,gradient elution from 0 to 40% ethyl acetate in hexanes) afforded(S)-4-bromo-2-(oxiran-2-ylmethoxy)phenyl acetate (0.72 g, 2.508 mmol) asa colorless liquid. LC/MS M⁺²³=309, 311.

Preparation 1C:(R)-(6-bromo-2,3-dihydrobenzo[b][1,4]dioxin-2-yl)methanol

(S)-4-bromo-2-(oxiran-2-ylmethoxy)phenyl acetate (0.72 g, 2.508 mmol)was dissolved in tetrahydrofuran (15 mL) and 2M aqueous solution ofsodium hydroxide (1.442 mL, 2.88 mmol) was added. The mixture wasvigorously stirred at room temperature for 2.5 days. Hexanes (7 mL) wasadded. The aqueous layer was separated and extracted with ethyl acetate(3×2 mL). The combined organic solutions were dried over anhydroussodium sulfate and concentrated. Flash chromatography purification (40 gsilica gel column, gradient elution from 0 to 40% ethyl acetate inhexanes) afforded(R)-(6-bromo-2,3-dihydrobenzo[b][1,4]dioxin-2-yl)methanol (0.54 g, 2.203mmol) as a white solid. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.04 (d, J=2.4Hz, 1H), 6.96 (dd, J=8.7, 2.3 Hz, 1H), 6.77 (d, J=8.6 Hz, 1H), 4.30 (dd,J=11.3, 2.3 Hz, 1H), 4.27-4.21 (m, 1H), 4.10 (dd, J=11.3, 7.7 Hz, 1H),3.95-3.80 (m, 2H).

Preparation 1D:(R)-6-bromo-2-((pentyloxy)methyl)-2,3-dihydrobenzo[b][1,4]dioxine

To a stirred solution of(R)-(6-bromo-2,3-dihydrobenzo[b][1,4]dioxin-2-yl) methanol (0.38 g,1.551 mmol) in anhydrous tetrahydrofuran (15 mL) was added sodiumhydride (60% mineral oil dispersion, 0.310 g, 7.75 mmol) portionwise atroom temperature under nitrogen. The resulting mixture was stirred atroom temperature for 15 min before n-amyl iodide (1.017 mL, 7.75 mmol)was added. The mixture was stirred at room temperature for 2 days.Saturated aqueous ammonium chloride solution (4 mL) and hexanes (10 mL)were added. The aqueous layer was separated and extracted with ethylacetate (2×3 mL). The combined organic solutions were dried overanhydrous sodium sulfate and concentrated. Flash chromatographypurification (12 g silica gel column, gradient elution from 0 to 20%ethyl acetate in hexanes) afforded(R)-6-bromo-2-((pentyloxy)methyl)-2,3-dihydrobenzo [b][1,4]dioxine (0.39g, 1.237 mmol) as a colorless liquid. ¹H NMR (400 MHz, CHLOROFORM-d) δ7.01 (d, J=2.4 Hz, 1H), 6.92 (dd, J=8.6, 2.2 Hz, 1H), 6.74 (d, J=8.6 Hz,1H), 4.31-4.23 (m, 2H), 4.07-3.98 (m, 1H), 3.71-3.63 (m, 1H), 3.58 (dd,J=10.3, 5.9 Hz, 1H), 3.48 (t, J=6.6 Hz, 2H), 1.64-1.51 (m, 2H),1.37-1.27 (m, 4H), 0.93-0.86 (m, 3H).

Preparation 1E: Ethyl1-((diphenylmethylene)amino)-4-((R)-2-((pentyloxy)methyl)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)cyclopent-2-enecarboxylate

An oven dried microwave vial with stir bar was charged with(R)-6-bromo-2-((pentyloxy)methyl)-2,3-dihydrobenzo[b][1,4]dioxine (390mg, 1.237 mmol), ethyl1-((diphenylmethylene)amino)cyclopent-3-enecarboxylate (593 mg, 1.856mmol), triphenylphosphine (64.9 mg, 0.247 mmol), palladium(II) acetate(27.8 mg, 0.124 mmol), potassium acetate (243 mg, 2.475 mmol), and DMA(3 mL). The mixture was sparged with nitrogen for 3 minutes. The mixturewas processed on a Personal Chemistry microwave (60 minutes at 140° C.).The mixture was mixed with water (60 mL) and extracted with ethylacetate (5×5 mL). The combined ethyl acetate extracts were dried overanhydrous sodium sulfate and concentrated under reduced pressure. Flashchromatography purification (40 g silica gel column, gradient elutionfrom 5 to 100% ethyl acetate in hexanes) afforded ethyl1-((diphenylmethylene)amino)-4-((R)-2-((pentyloxy)methyl)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)cyclopent-2-enecarboxylate(300 mg, 0.542 mmol) as a sticky liquid. LC/MS M⁺¹=554.

Example 1

To a stirred solution of ethyl1-((diphenylmethylene)amino)-4-((R)-2-((pentyloxy)methyl)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)cyclopent-2-enecarboxylate(300 mg, 0.542 mmol) and water (0.27 mL) in diethyl ether (8 mL) wasadded 6 N aqueous hydrochloric acid (0.542 mL, 3.25 mmol). The mixturewas stirred at room temperature for 30 min and then basified withpotassium carbonate solid and water (1 mL). The mixture was extractedwith ethyl acetate (4×4 mL). The combined ethyl acetate extracts weredried over anhydrous sodium sulfate and concentrated under reducedpressure to give a liquid. The liquid was dissolved in EtOH (10 mL).Sodium borohydride (123 mg, 3.25 mmol) was added. The mixture wasstirred at room temperature overnight. Next, 6N aqueous hydrochloricacid (2 mL) was added slowly to make pH approximately 2. The mixture wasstirred at room temperature for 1 h. The mixture was basified to pHapproximately 12 with 2 N aqueous sodium hydroxide solution. Afterstirring at room temperature for 30 min, the mixture was concentrated.The aqueous residue was extracted with ethyl acetate (4×4 mL). Thecombined ethyl acetate extracts were dried over anhydrous sodium sulfateand concentrated under reduced pressure to give a solid.

The solid material was dissolved in MeOH (10 mL) and acetic acid (1 mL).Next, 10% Pd/C (100 mg, 0.094 mmol) was added under nitrogen. Themixture was hydrogenated under hydrogen balloon overnight. The catalystwas filtered and washed with methanol. The filtrate was concentrated.The residue was mixed with water (3 mL), basified with potassiumcarbonate solid, and extracted with ethyl acetate (5×3 mL). The combinedethyl acetate extracts were dried over anhydrous sodium sulfate andconcentrated. Flash chromatography purification (4 g silica gel column,gradient elution from 0->20% of 2M ammonia in methanol solution inEtOAc) afforded(1-amino-3-((R)-2-((pentyloxy)methyl)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)cyclopentyl)methanol(180 mg, 0.515 mmol) as a semisolid. LC/MS M⁺¹=350.

The semisolid was separated into three fractions using chiral SFC(Cell-4 (25×3 cm, 5 μm), CO₂/(MeOH+0.5% DEA)=60/40, 130 ml/min, 284 nm,35° C.). Fractions 1 and 3 were individually concentrated and purifiedusing reverse phase HPLC (Phenomenex Luna 5u 30×100 mm (Axia), solventA: 10% MeOH: 90% H₂O: 0.1% TFA, solvent B: 90% MeOH, 10% H₂O, 0.1% TFA).Concentration, basification with potassium carbonate, and extractionwith ethyl acetate gave the corresponding compounds. Fraction 2 wasconcentrated and separated using chiral SFC (Cell-2-H (3×25 cm), 40% IPAw 0.1% DEA and 0.1% water in CO₂, 150 ml/min, 220 nm, 50° C.) to giveFraction 2-A and Fraction 2-B as glassy solids. All four isomers havethe same molecular weights. LC/MS M⁺¹=350.

Example 1(Fraction 1): ¹H NMR (400 MHz, METHANOL-d₄) δ 6.77 (d, J=0.9Hz, 1H), 6.75-6.73 (m, 2H), 4.30-4.21 (m, 2H), 4.04-3.96 (m, 1H),3.71-3.57 (m, 2H), 3.53-3.40 (m, 4H), 2.97 (tt, J=11.3, 7.1 Hz, 1H),2.19 (dd, J=13.1, 7.6 Hz, 1H), 2.04-1.63 (m, 4H), 1.63-1.54 (m, 2H),1.54-1.43 (m, 1H), 1.40-1.29 (m, 4H), 0.96-0.87 (m, 3H).

Example 2 (Fraction 3): ¹H NMR (400 MHz, METHANOL-d₄) δ 6.77 (d, J=0.9Hz, 1H), 6.75-6.73 (m, 2H), 4.29-4.21 (m, 2H), 4.03-3.95 (m, 1H),3.70-3.57 (m, 2H), 3.54-3.40 (m, 4H), 2.97 (tt, J=11.3, 7.1 Hz, 1H),2.19 (dd, J=12.8, 7.3 Hz, 1H), 2.04-1.63 (m, 4H), 1.63-1.54 (m, 2H),1.54-1.45 (m, 1H), 1.39-1.30 (m, 4H), 0.95-0.87 (m, 3H).

Example 3 (Fraction 2-A): ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.77-6.68 (m,3H), 4.29-4.20 (m, 2H), 4.03-3.96 (m, 1H), 3.71-3.55 (m, 2H), 3.53-3.44(m, 4H), 3.26-3.17 (m, 1H), 2.16-2.05 (m, 1H), 2.03-1.95 (m, 1H),1.93-1.85 (m, 1H), 1.74-1.53 (m, 4H), 1.41-1.23 (m, 5H), 0.95-0.88 (m,3H).

Example 4 (Fraction 2-B): ¹H NMR (400 MHz, METHANOL-d₄) δ 6.78-6.68 (m,3H), 4.29-4.21 (m, 2H), 4.04-3.96 (m, 1H), 3.71-3.57 (m, 2H), 3.56-3.46(m, 4H), 3.22 (q, J=7.3 Hz, 1H), 2.17-1.87 (m, 3H), 1.75-1.53 (m, 4H),1.41-1.23 (m, 5H), 0.97-0.86 (m, 3H).

Using the general procedures for the preparation of Examples 1 to 4, thefollowing compounds were prepared from the corresponding aryl bromideintermediates. The compounds were analyzed using HPLC condition C.

TABLE 1 HPLC Ex. ret. time MS No. Structure MW (min.) (M + 1) Comment  5 6

335.4 335.4 2.50 2.49 336 336 Cis- cyclopentyl Isomer 1 Cis- cyclopentylIsomer 2  7  8  9 10

344.5 344.5 344.5 344.5 2.61 2.63 2.61 2.63 345 345 345 345 Isomer 1Isomer 2 Isomer 3 Isomer 4 11 12 13 14

331.5 331.5 331.5 331.5 3.09 3.08 3.05 3.05 332 332 332 332 Cyclohexaneisomer 1 Mixture of 2 trans cyclopentyl isomers Cyclohexane isomer 1Mixture of 2 cis cyclopentyl isomers Cyclohexane isomer 2 Mixture of 2trans cyclopentyl isomers Cyclohexane isomer 2 Mixture of 2 ciscyclopentyl isomers 15 16

331.5 331.5 3.38 3.36 332 332 pyran isomer 1 cis-cyclopentyl isomer 1pyran isomer 1 cis-cyclopentyl isomer 2 17 18

331.5 331.5 3.35 3.36 332 332 pyran isomer 2 cis- cyclopentyl isomer 1pyran isomer 2 cis- cyclopentyl isomer 2 19

331.5 3.31 332 Mixture of 2 diastereomers 20 21 22 23

331.5 331.5 331.5 331.5 3.41 3.51 3.38 3.52 332 332 332 332 pyran isomer1 cis-cyclopentyl isomer 1 pyran isomer 1 cis-cyclopentyl isomer 2 pyranisomer 2 cis-cyclopentyl isomer 1 pyran isomer 2 cis-cyclopentyl isomer1 24

331.5 3.22 332 25

331.5 3.23 332 26

345.5 3.43 346 27

345.5 3.52 346

Examples 28 and 29((1R,3S)-1-amino-3-(6-(pentyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 28A:(5R,7S)-7-(6-(pentyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

A mixture of 1-pentanol (10 mL, 92 mmol), p-toluenesulfonic acidmonohydrate (8.00 mg, 0.042 mmol), and trimethoxymethane (0.613 mL, 5.61mmol) was stirred at 100° C. for 2 h with a slow nitrogen stream toremove methanol byproduct. The residual liquid was mixed with(5R,7S)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(400 mg, 1.402 mmol) and stirred at 100° C. under nitrogen for 2.5 h.Next, 10% Pd/C (400 mg) was added at room temperature, followed by ethylacetate (5 mL). The mixture was vigorously stirred under hydrogenballoon for 4 h. The mixture was filtered through a membrane filter andthe filtrate was concentrated. Flash chromatography purification (12 gsilica gel column, gradient elution from 0 to 100% ethyl acetate inhexanes) afforded(5R,7S)-7-(6-(pentyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(350 mg, 0.979 mmol) as a sticky solid. LC/MS M⁺¹=358. Chiral separation(Lux-Amy-2 (3×25 cm), 25% MeOH, 120 ml/min, 220 nm, 45° C.) of the solidgave two isomers. Each isomer was hydrolyzed in the following fashion.

Example 28 (Isomer 1)

A mixture of(5R,7S)-7-(6-(pentyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(110 mg, 0.308 mmol), lithium hydroxide monohydrate (155 mg, 3.69 mmol),dioxane (1 mL), and water (1 mL) was stirred at 90° C. under nitrogenfor 15 h. The mixture was cooled and extracted with ethyl acetate (4×1mL). The combined ethyl acetate extracts were dried over anhydroussodium sulfate and concentrated under reduced pressure. Purificationusing reverse phase HPLC (Phenomenex Luna 5u 30×100 mm (Axia); gradientover 8 min from 30 to 100% of solvent B; solvent A: 10% MeOH: 90% H₂O:0.1% TFA; solvent B: 90% MeOH, 10% H₂O, 0.1% TFA), concentration,basification with potassium carbonate, and extraction with ethyl acetategave((1R,3S)-1-amino-3-(6-(pentyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol (61 mg, 0.173 mmol,) as a white solid. LC/MS M⁺¹=332. ¹H NMR(400 MHz, CHLOROFORM-d) δ 7.03-6.95 (m, 3H), 3.73-3.63 (m, 1H),3.57-3.42 (m, 4H), 3.09-2.97 (m, 2H), 2.95-2.85 (m, 1H), 2.82-2.68 (m,2H), 2.27 (dd, J=13.3, 7.8 Hz, 1H), 2.13-2.01 (m, 2H), 1.97-1.46 (m,7H), 1.37-1.29 (m, 4H), 0.94-0.87 (m, 3H).

Example 29 (Isomer 2)

¹H NMR (400 MHz, CHLOROFORM-d) δ 7.03-6.95 (m, 3H), 3.73-3.64 (m, 1H),3.57-3.49 (m, 2H), 3.48-3.40 (m, 2H), 3.09-2.96 (m, 2H), 2.95-2.86 (m,1H), 2.81-2.69 (m, 2H), 2.25 (dd, J=13.2, 7.9 Hz, 1H), 2.13-2.00 (m,2H), 1.95-1.83 (m, 1H), 1.82-1.55 (m, 5H), 1.48 (dd, J=13.2, 11.0 Hz,1H), 1.37-1.29 (m, 4H), 0.93-0.87 (m, 3H).

Examples 30 and 31((1R,3S)-1-amino-3-(6-(heptyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 30A:(5R,7S)-7-(6-(heptyloxy)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of(5R,7S)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(100 mg, 0.350 mmol) and 1-heptanol (500 μL, 3.54 mmol) in toluene (2mL) was added p-toluenesulfonic acid monohydrate (5 mg, 0.026 mmol).

Oven dried 3-angstrom molecular sieves were added and the mixture washeated at reflux overnight. The reaction mixture was diluted with ethylacetate and washed with saturated NaCl. The organic layer was dried withMgSO₄, filtered, and concentrated. The crude material was purified on asilica gel cartridge (40 g) using an EtOAc/Hex gradient (0-100% EtOAcover 20 minutes) to afford 55 mg of(5R,7S)-7-(6-(heptyloxy)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.HPLC retention time=1.26 min (condition G); LC/MS M⁺¹=384. ¹H NMR (400MHz, CHLOROFORM-d) δ 7.00-6.86 (m, 3H), 5.79 (s, 1H), 5.52 (s, 1H),4.41-4.24 (m, 2H), 3.86 (t, J=6.6 Hz, 2H), 2.94-2.82 (m, 2H), 2.41 (t,J=8.0 Hz, 2H), 2.36-2.25 (m, 1H), 2.20-2.07 (m, 2H), 2.05-1.90 (m, 2H),1.79-1.70 (m, 2H), 1.51-1.21 (m, 10H), 0.98-0.83 (m, 3H).

Preparation 30B:(5R,7S)-7-(6-(pentyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of(5R,7S)-7-(6-(heptyloxy)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(53 mg, 0.138 mmol) in MeOH (10 mL) was added Pearlman's Catalyst (19.41mg, 0.138 mmol). The reaction mixture was hydrogenated under a balloonof H₂ for 2 hours. The catalyst was filtered away, and then concentratedin vacuo. The individual isomers were separated using a CHIRALPAK® AS-Hcolumn under SFC conditions (30% MeOH in CO₂).

Isomer 1 (30-B-i, 9 mg) Chiral HPLC retention time=8.55 min; LC/MSM⁺¹=386. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.10-7.03 (m, 1H), 7.00-6.91(m, 2H), 5.29 (br. s., 1H), 4.40-4.26 (m, 2H), 3.78-3.67 (m, 1H), 3.55(qd, J=6.6, 2.4 Hz, 2H), 3.13-2.98 (m, 2H), 2.98-2.88 (m, 1H), 2.84-2.71(m, 2H), 2.33 (dd, J=13.2, 7.3 Hz, 1H), 2.23-2.04 (m, 3H), 1.96 (dd,J=13.1, 10.9 Hz, 2H), 1.88-1.74 (m, 2H), 1.61 (quin, J=6.9 Hz, 4H),1.43-1.21 (m, 6H), 0.95-0.85 (m, 3H).

Isomer 2 (30-B-ii, 9 mg) Chiral HPLC retention time=9.81 min; LC/MSM⁺¹=386. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.09-7.03 (m, 1H), 7.00-6.90(m, 2H), 5.25 (s, 1H), 4.40-4.24 (m, 2H), 3.80-3.67 (m, 1H), 3.55 (qd,J=6.6, 2.4 Hz, 2H), 3.14-2.99 (m, 2H), 2.98-2.87 (m, 1H), 2.84-2.69 (m,2H), 2.33 (dd, J=13.2, 7.3 Hz, 1H), 2.22-2.04 (m, 3H), 2.02-1.90 (m,2H), 1.88-1.74 (m, 2H), 1.60 (q, J=7.0 Hz, 4H), 1.43-1.20 (m, 6H),0.95-0.85 (m, 3H). The absolute stereochemistry of the isomers was notdetermined.

Example 30:((1R,3S)-1-amino-3-(6-(heptyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a mixture of(5R,7S)-7-(6-(heptyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one,Isomer 1 (30B-i, 9 mg, 0.023 mmol) in dioxane (4 mL) was added 1N NaOH.The reaction mixture was heated at 100° C. overnight, and then cooledand acidified with TFA. The mixture was concentrated in vacuo, thentriturated with MeOH, and filtered. The filtrate was purified filtrateby HPLC. HPLC conditions: Phenomenex Luna 5 micron C18 column (30×100mm); MeCN (0.1% TFA)/water (0.1% TFA); 20%-100% gradient over 15minutes; 30 mL/min. Isolated fractions with correct mass andfreeze-dried overnight. Recovered 5 mg of((1R,3S)-1-amino-3-(6-(heptyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(Example 30): HPLC retention time=9.31 min (condition H); LC/MS M⁺¹=360.¹H NMR (400 MHz, METHANOL-d₄) δ 7.07-6.98 (m, 3H), 3.83-3.73 (m, 1H),3.71-3.51 (m, 4H), 3.18-3.08 (m, 1H), 3.04 (dd, J=16.6, 5.0 Hz, 1H),2.96-2.85 (m, 1H), 2.82-2.68 (m, 2H), 2.42 (ddd, J=13.3, 7.1, 1.2 Hz,1H), 2.17-2.01 (m, 2H), 2.00-1.89 (m, 3H), 1.88-1.77 (m, 1H), 1.73 (t,J=12.8 Hz, 1H), 1.59 (quin, J=6.9 Hz, 2H), 1.45-1.22 (m, 8H), 0.96-0.86(m, 3H).

Example 31:((1R,3S)-1-amino-3-(6-(heptyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a mixture of(5R,7S)-7-(6-(heptyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-oneIsomer 2 (30-B-ii, 8 mg, 0.021 mmol) in dioxane (4 mL) was added 1NNaOH. The mixture was heated at 100° C. overnight, and then cooled andacidified with TFA. The mixture was concentrated in vacuo thentriturated with MeOH, and filtered. The filtrate was purified by HPLC.HPLC conditions: Phenomenex Luna 5 micron C18 column (30×100 mm); MeCN(0.1% TFA)/water (0.1% TFA); 20%-100% gradient over 15 minutes; 30mL/min. Isolated fractions with correct mass and freeze-dried overnight.Recovered 5 mg of((1R,3S)-1-amino-3-(6-(heptyloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(Example 31). HPLC retention time=9.34 min (condition H); LC/MS M⁺¹=360.¹H NMR (400 MHz, METHANOL-d₄) δ 7.03 (s, 2H), 7.00 (s, 1H), 3.83-3.73(m, 1H), 3.70-3.61 (m, 2H), 3.61-3.50 (m, 2H), 3.18-3.08 (m, 1H), 3.04(dd, J=16.4, 4.7 Hz, 1H), 2.97-2.84 (m, 1H), 2.81-2.66 (m, 2H), 2.42(ddd, J=13.4, 7.1, 1.1 Hz, 1H), 2.19-2.01 (m, 2H), 2.00-1.89 (m, 3H),1.88-1.77 (m, 1H), 1.73 (t, J=12.8 Hz, 1H), 1.59 (quin, J=6.9 Hz, 2H),1.44-1.23 (m, 8H), 0.97-0.87 (m, 3H).

The following compounds were prepared according to the generalprocedures of Examples 28 and 29.

TABLE 2 Ex. HPLC HPLC No. Structure MW ret. time (min.) condition MS(M⁺¹) Comment 32 33

331.5 331.5 3.23 3.23 K K 332 332 Isomer 1 Isomer 2 34 35

345.5 345.5 3.29 3.26 C C 346 346 Isomer 1 Isomer 2 36 37

317.5 317.5 7.59 7.59 H H 318 318 Isomer 1 Isomer 2

Example 38((1R,3S)-1-amino-3-((S)-6-((Z)-hex-2-en-1-yloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 38A:2-(((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)oxy)acetaldehyde

A solution of oxalyl dichloride (68.9 mg, 0.543 mmol) in DCM (3 ml) wasstirred under N₂ and cooled to −78° C. DMSO (64.2 μl, 0.905 mmol) wasthen added dropwise and stirred for 1 h at the temperature, a solutionof(5R,7S)-7-((S)-6-(2-hydroxyethoxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(150 mg, 0.453 mmol) in DCM (3 mL) (several drops of DMSO was added tohelp dissolve the compound) was added dropwise and the mixture wasstirred for 30 min. at −78° C. Then TEA (252 μl, 1.810 mmol) was addeddropwise and the mixture was stirred for 15 min at −78° C. and warmed toroom temperature and stirred for 15 min. The mixture was quenched withwater (1 mL) at 0° C., diluted with EtOAc (50 mL), washed with saturatedNH₄Cl (2×30 mL), dried (Na₂SO₄) and concentrated under vacuo to give2-(((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)oxy)acetaldehyde,(150 mg) as a white solid. LC/MS M⁺¹=330.

Preparation 38B (Isomer 1) (Condition 1):(5R,7S)-7-((S)-6-((Z)-hex-2-en-1-yloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of butyltriphenylphosphonium bromide (116 mg, 0.364 mmol)in THF (3 mL) at −78° C. and under nitrogen was slowly added n-butyllithium in hexane (239 μl, 0.383 mmol). The solution was stirred at −78°C. for 15 min, and then stirred at 0° C. for 30 min (light orangecolor). To the solution was added2-(((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)oxy)acetaldehyde(60 mg, 0.182 mmol) in THF (3 mL) at −78° C. The reaction mixture wasstirred at −78° C. for 15 min and stirred at room temperature for 2 h.The reaction was quenched with saturated aqueous ammonium chloride andextracted with ethyl acetate. The organic extract was washed withsaturated NH₄Cl (3×20 mL), dried over sodium sulfate, filtered, andconcentrated in vacuo to give the desired product as white solid,(5R,7S)-7-((S)-6-((Z)-hex-2-en-1-yloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one,LC/MS M⁺¹=370.

Preparation 38B (Isomer 2) (Condition 2):(5R,7S)-7-((S)-6-((E)-hex-2-en-1-yloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

KHMDS (683 μl, 0.683 mmol) was added dropwise to a solution of5-(butylsulfonyl)-1-phenyl-1H-tetrazole (80 mg, 0.301 mmol) and2-(((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)oxy)acetaldehyde (90 mg, 0.273 mmol) in THF (15 ml) at −78° C. The resultantsolution was stirred at the temperature for 2 h and warmed to roomtemperature and stirred for 16 h.

Next, water (1 ml) was added with acetone-dry ice cooling, and themixture was warmed to room temperature, followed by the addition ofwater (10 ml), extracted with EtOAc (30 ml), washed with saturatedNaHCO₃(2×15 ml), brine (20 ml), dried (Na₂SO₄) and concentrated undervacuo to give the desired product which was purified with flashchromatography using ISCO column (12 g, EtOAc/Hexane=0%-40%,), to give(5R,7S)-7-((S)-6-((E)-hex-2-en-1-yloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one,15 mg, LC/MS M⁺¹=370.

Example 38

To a solution of(5R,7S)-7-((S)-6-((Z)-hex-2-en-1-yloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(90 mg, 0.244 mmol) in dioxane (2 mL) was added lithium hydroxide (58.3mg, 2.436 mmol) in water (1 mL) and stirred at 100° C. for 16 h. Thereaction mixture was diluted with water and extracted with EtOAc. Theorganic layer was collected, dried over Na₂SO₄, concentrated on therotavapor to give the crude product which was purified with preparativeHPLC: column Phenomenex Luna C18 5u 21.2×100 mm. Solvent A: 10% MeOH-90%H₂O-0.1% TFA; Solvent B: 90% MeOH-10% H₂O-0.1% TFA. Gradient time=15min. Start B=0%, Final B 100%. Stop time 25 min. to afford((1R,3S)-1-amino-3-((S)-6-((Z)-hex-2-en-1-yloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,LC/MS M⁺¹=344. HPLC retention time=8.20 min. (Condition L), ¹H NMR (400MHz, METHANOL-d₄) δ 7.10-6.90 (m, 3H), 5.70-5.60 (m, 2H), 4.2 (m, 2H),3.8 (m, 1H), 3.65 (m, 2H), 3.25-2.72 (m, 5H), 2.40 (m, 1H), 2.15 (m,3H), 2.10-1.72 (m, 6H), 1.44 (m, 3H), 0.92 (t, J=7.5 Hz, 3H).

Using the general procedure of Example 38, the following compounds wereprepared.

TABLE 3 HPLC Ex. ret. time HPLC No. Structure MW (min.) condition MS(M⁺¹) Comment 39

343.5 8.21 L 344 Step B Condition 1 40

343.5 7.23 L 344 Step B Condition 2 41

343.5 7.25 L 344 Step B Condition 2 42

357.5 7.62 L 358 Step B Condition 1

Example 43((1R,3S)-1-amino-3-((R)-6-((4-ethylbenzyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 43A:(R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carbaldehyde

A solution of oxalyl chloride (261 μl, 2.99 mmol) in DCM (5 ml) wasstirred under N₂ and cooled to −78° C. DMSO (424 μl, 5.97 mmol) was thenadded dropwise and stirred for 1 h at the temperature, a solution of(5R,7S)-7-((R)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(Preparation 51C, 600 mg, 1.991 mmol) in DCM (3 ml)/1 ml of DMSO wasadded dropwise and the mixture was stirred for 30 min. at the sametemperature. Then TEA (1110 μl, 7.96 mmol) was added dropwise and themixture was stirred for 15 min and warmed to room temperature andstirred for another 15 min. The mixture was quenched with water (1 ml)at 0° C., diluted with EtOAc (50 ml), washed with saturated NH₄Cl (2×30ml), dried (Na₂SO₄), and concentrated under vacuo. The residue waspurified with flash chromatography (25 g, EtOAc/Hexane=0-100%, gradienttime=15 min) to recover 500 mg desired product(R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carbaldehyde(500 mg). LC/MS M⁺¹=300.

Preparation 43B:(5R,7S)-7-((R)-6-((S)-1-hydroxyethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a stirred mixture of(R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carbaldehyde(500 mg, 1.670 mmol) and THF (5 mL) was added a solution ofmethylmagnesium bromide (2227 μl, 6.68 mmol) (3M in diethyl ether)dropwise at −78° C. The solution was gradually warmed up to roomtemperature and stirred overnight under nitrogen. The reaction wasquenched with water at 0° C. The mixture was extracted with EtOAc (30ml), washed with saturated NH₄Cl (2×30 ml), brine (20 ml), dried(Na₂SO₄) and purified with flash chromatography (25 g,EtOAc/Hexane=0-100%, gradient time=12.5 min) to recover(5R,7S)-7-((R)-6-((S)-1-hydroxyethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(460 mg). LC/MS M⁺¹=316.

Preparation 43C:(5R,7S)-7-((R)-6-acetyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

A solution of oxalyl chloride (511 μl, 5.83 mmol) in DCM (5 ml) wasstirred under N₂ and cooled to −78° C. DMSO (621 μl, 8.75 mmol) was thenadded dropwise and the mixture was stirred for 1 h at −78° C. A solutionof(5R,7S)-7-((R)-6-((S)-1-hydroxyethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(460 mg, 1.458 mmol) in DCM (3 ml) was added dropwise and the mixturewas stirred for 30 min. at −78° C. Then TEA (1220 μl, 8.75 mmol) wasadded dropwise and the mixture was stirred for 15 min and warmed to roomtemperature and stirred for 15 min. The mixture was quenched with water(1 ml) at 0° C., diluted with EtOAc (50 ml), which was washed withsaturated NH₄Cl (2×30 ml), dried (Na₂SO₄) and concentrated under vacuo.The residue was purified with flash chromatography (25 g,EtOAc/hexane=0-100%, gradient time=15 min) to recover the desiredcompound(5R,7S)-7-((R)-6-acetyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(239 mg). LC/MS M⁺¹=314.

Preparation 43D:(R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-ylacetate

To a solution of(5R,7S)-7-((R)-6-acetyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(220 mg, 0.702 mmol) in DCM (4 ml) was added 77% m-CPBA (315 mg, 1.404mmol) in portions. The reaction mixture was stirred at room temperaturefor 60 h before it was washed with 0.2 N aqueous NaOH (10 ml). The washsolution was extracted back with DCM (2×15 ml). The combined organicextracts were dried over Na₂SO₄ and the solvent was removed in vacuo.The residue was purified with flash chromatography (12 g,EtOAc/Hexane=0-60%, gradient time=15 min) to recover the desired product(R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-ylacetate (220 mg). LC/MS M⁺¹=330.

Preparation 43E:(5R,7S)-7-((R)-6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To the solution of(R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-ylacetate (200 mg, 0.607 mmol) in MeOH (2 ml), sodium hydroxide (1822 μl,1.822 mmol) was added and the mixture was stirred at room temperaturefor 1 h. The mixture was taken in EtOAc (20 ml), washed with saturatedNaHCO₃(10 ml) and brine (10 ml), dried (Na₂SO₄) and concentrated undervacuo to get the desired product which was used to next step as was(5R,7S)-7-((R)-6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(165 mg). LC/MS M⁺¹=288.

Preparation 43F:(5R,7S)-7-((R)-6-((4-ethylbenzyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

The(5R,7S)-7-((R)-6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(15 mg, 0.052 mmol) was dissolved in anhydrous nitromethane (1.5 ml) ina dry 8 ml tube. Anhydrous iron (III) chloride (2 mg, 0.012 mmol),4-ethylbenzaldehyde (14.01 mg, 0.104 mmol) and triethylsilane (12.14 mg,0.104 mmol) were added and the resulting solution was stirred at roomtemperature for 2 h. Next, 10 ml of water was added and the aqueouslayer was extracted with DCM (2×15 ml). The combined organic layers werewashed with brine and dried (Na₂SO₄). The mixture was filtered andconcentrated. The residue was purified via gradient flash chromatography(0-60% EtOAc in hexanes, ISCO column 12 g) which provided of the desiredproduct(5R,7S)-7-((R)-6-((4-ethylbenzyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(18 mg). LC/MS M⁺¹=406.

Example 43

(5R,7S)-7-((R)-6-((4-ethylbenzyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(20 mg, 0.049 mmol) was mixed with lithium hydroxide hydrate (31.0 mg,0.740 mmol) in 1,4-dioxane (2 ml) and water (0.5 ml), the mixture wasstirred at 100° C. overnight under N₂. The solution was concentratedunder vacuo and the residue was dissolved in DCM (40 ml), washed withwater (15 ml) and brine (10 ml), dried (Na₂SO₄) and concentrated. Thesolid was mixed with MeCN (2 ml), the solvent was removed, and the solidwas dried under vacuum overnight to give the crude product which waspurified with preparative HPLC. Phenomenex Luna C 18 5u (21.2×150 mm),Solvent A: 10% MeOH-90% H₂O-0.1% TFA; Solvent B: 90% MeOH-10% H₂O-0.1%TFA, Start B %=0, Final % B=100. Gradient time 15 min, stop time 20 min.to afford((1R,3S)-1-amino-3-((R)-6-((4-ethylbenzyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl) methanol (15 mg). LC/MS M⁺¹=380. HPLC Rt=7.61 (conditionL). ¹H NMR (400 MHz, METHANOL-d₄) δ 7.35-7.25 (m, 2H), 7.23-7.15 (m,2H), 7.06-6.94 (m, 3H), 4.66-4.55 (m, 2H), 3.94-3.83 (m, 1H), 3.57-3.41(m, 2H), 3.12-2.88 (m, 3H), 2.84-2.59 (m, 4H), 2.26-1.66 (m, 7H),1.59-1.46 (m, 1H), 1.29-1.17 (m, 3H).

The examples in Table 4 were prepared according to the general procedureof Example 43.

TABLE 4 HPLC Ex. ret. time HPLC MS No. Structure MW (min.) condition(M⁺¹) 44

351.49 6.70 L 352 45

365.51 8.17 L 366 46

381.51 6.82 L 382 47

381.51 6.82 L 382 48

395.54 7.10 L 396 49

435.48 8.82 L 436

Examples 50 and 51((1R,3S)-1-amino-3-(6-((benzyloxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 50A: Methyl4-oxo-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate

To a mixture of potassium carbonate (523 mg, 3.78 mmol),(5R,7S)-7-(4-bromophenyl)-3-oxa-1-azaspiro[4.4]nonan-2-one (800 mg, 2.70mmol), itaconic acid (457 mg, 3.51 mmol), and acetonitrile (8 mL) wasadded water (2.4 mL). The mixture was stirred till the evolution ofcarbon dioxide stopped and then bubbled with nitrogen for 3 min. Afterpalladium(II) acetate (30.3 mg, 0.135 mmol) and tri-o-tolylphosphine (82mg, 0.270 mmol) were added, the mixture was bubbled with nitrogen for anadditional 3 min. The mixture was stirred at 80° C. for 20 h and thenconcentrated. The residue was mixed with water (40 mL), basified withpotassium carbonate and filtered. The filtrate was washed with diethylether (2×15), acidified to pH approximately 2 with 6N aqueoushydrochloric acid. The solid was separated and the aqueous solution wasextracted with a mixture of THF/EtOAc (3:1) (4×10 mL). The solid and theextracts were combined and concentrated. LC/MS [M−H₂O]⁺¹=328.

The residue was mixed with THF (5 mL), ethyl acetate (5 mL), methanol(20 mL), and 10% Pd/C (400 mg, 0.376 mmol) and hydrogenated underhydrogen balloon overnight. The catalyst was filtered off through amembrane filter and washed with methanol. The filtrate was concentratedand lyophilized to give a solid. LC/MS [M−H₂O]⁺¹=330.

The solid was mixed with 98% sulfuric acid (15 mL, 281 mmol). The clearsolution was stirred at room temperature for 4 h. Methanol (8 mL, 198mmol) was added slowly with water-bath cooling. The mixture was stirredat room temperature for 1 h before being poured onto ice (150 g). Themixture was extracted with ethyl acetate (4×40 mL). The combined ethylacetate extracts were washed with saturated aqueous sodium bicarbonatesolution (20 mL), dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. Flash chromatography purification (24 g silicagel column, gradient elution from 10 to 100% ethyl acetate in hexanes)afforded methyl4-oxo-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(440 mg, 1.281 mmol). LC/MS M⁺¹=344.

Preparation 50B: Methyl6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate

A mixture of methyl4-oxo-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(440 mg, 1.281 mmol), MeOH (15 mL), acetic acid (1.5 mL), and 10% Pd/C(200 mg, 0.188 mmol) was hydrogenated under hydrogen balloon over aperiod of two days. The mixture was filtered through a membrane filter.The filtrate was concentrated to give methyl6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(337 mg, 1.023 mmol) as a white solid. LC/MS M⁺¹=330.

Preparations 50C and 51C:(5R,7S)-7-(6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

A mixture of methyl6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(337 mg, 1.023 mmol), anhydrous tetrahydrofuran (3 mL), and 2N THFsolution of lithium borohydride (2.56 mL, 5.12 mmol) was stirred at 70°C. for 4 h. Saturated aqueous ammonium chloride solution was addedslowly at 0° C. to quench the reaction. Water and ethyl acetate wereadded. The aqueous solution was extracted with ethyl acetate. Thecombined ethyl acetate solutions were dried over anhydrous sodiumsulfate and concentrated under reduced pressure to give(5R,7S)-7-(6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(300 mg, 0.995 mmol). LC/MS M⁺¹=302. Chiral SFC separation (AD-H(0.46×25 cm), 45% MeOH in C02, 3 ml/min, 220 nm, 35° C.) gaveenantiomers 50C and 51C as white solids. Isomer 50C:(5R,7S)-7-((S)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.Isomer 51C:(5R,7S)-7-((R)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.Each enantiomer was independently converted to derivatives asillustrated below.

Preparation 51D:((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate

(5R,7S)-7-((R)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(enantiomer 51C; 690 mg, 2.289 mmol) was dissolved in dry pyridine (5mL) and p-toluenesulfonyl chloride (1309 mg, 6.87 mmol) was added in oneportion. The resulting mixture was reacted at room temperature for 4 h.The solvent was removed in vacuo. The residue was dissolved in methylenechloride and methanol. Flash chromatography purification (40 g silicagel column, gradient elution from 20 to 100% ethyl acetate in hexanes)afforded((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (860 mg, 1.888 mmol) as a white solid. LC/MSM⁺¹=456.

Example 51

To a stirred mixture of benzyl alcohol (0.031 mL, 0.296 mmol) and 1N THEsolution of potassium tert-butoxide (0.263 mL, 0.263 mmol) was added((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (15 mg, 0.033 mmol). The resulting mixture wasstirred at 70° C. under nitrogen overnight. The mixture wasconcentrated. The residue was mixed with water (0.5 mL), lithiumhydroxide monohydrate (28 mg, 0.66 mmol), and dioxane (1 mL). Theresulting mixture was stirred at 100° C. under nitrogen for 7 h and roomtemperature overnight. The mixture was extracted with ethyl acetate (4×1mL) and the combined ethyl acetate extracts were dried and concentrated.The crude material was purified via preparative LC/MS with the followingconditions: Column: Waters XBridge C18, 19×150 mm, 5-μm particles; GuardColumn: Waters XBridge C18, 19×10 mm, 5-m particles; Mobile Phase A:5:95 acetonitrile:water with 10-mM ammonium acetate; Mobile Phase B:95:5 acetonitrile:water with 10 mM ammonium acetate; Gradient: 15-100% Bover 15 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min.Fractions containing the desired product were combined and dried to give((1R,3S)-1-amino-3-((R)-6-((benzyloxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(8.0 mg, 0.022 mmol) as a solid. HPLC retention time=1.66 min (conditionA) LC/MS M⁺¹=366. ¹H NMR (500 MHz, METHANOL-d₄) δ 7.38-7.34 (m, 4H),7.33-7.27 (m, 1H), 7.03-6.96 (m, 3H), 4.56 (s, 2H), 3.56-3.44 (m, 4H),3.08-2.97 (m, 1H), 2.87 (dd, J=15.9, 4.5 Hz, 1H), 2.83-2.77 (m, 2H),2.47 (dd, J=16.3, 10.9 Hz, 1H), 2.26 (dd, J=12.9, 7.4 Hz, 1H), 2.16-1.98(m, 3H), 1.98-1.86 (m, 1H), 1.84-1.69 (m, 2H), 1.61-1.53 (m, 1H),1.51-1.40 (m, 1H).

Example 50

Example 50 was prepared from Isomer 50C:(5R,7S)-7-((S)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-oneusing the procedures of Step D and Step E. HPLC retention time=1.65 min(condition A); LC/MS M+1=366. ¹H NMR (500 MHz, METHANOL-d₄) δ 7.62 (s,1H), 7.34-7.38 (m, 3H), 7.30 (dq, J=8.8, 4.2 Hz, 1H), 7.04-6.96 (m, 3H),4.56 (s, 2H), 3.61-3.50 (m, 2H), 3.48 (d, J=6.9 Hz, 2H), 3.10-2.98 (m,1H), 2.88 (dd, J=16.3, 4.5 Hz, 1H), 2.83-2.76 (m, 2H), 2.47 (dd, J=16.3,10.4 Hz, 1H), 2.33 (dd, J=13.4, 7.4 Hz, 1H), 2.17-1.99 (m, 3H),1.99-1.79 (m, 3H), 1.66 (t, J=12.4 Hz, 1H), 1.51-1.39 (m, 1H).

The Examples in Table 5 were prepared according to the generalprocedures of Examples 50 and 51.

TABLE 5 HPLC ret. Ex. time HPLC MS No. Structure MW (min.) condition(M⁺¹) Comment 52 53

331.5 331.5 1.68 3.09 A A 332 332 Isomer 1 Isomer 2 54 55

345.5 345.5 1.85 1.85 A A 346 346 Isomer 1 Isomer 2 56 57

369.5 369.5 1.68 3.18 A A 370 370 Isomer 1 Isomer 2 58

347.5 1.06 A 348 Isomer 1 59

361.5 1.18 A 362 Isomer 1 60 61

347.5 347.5 1.07 1.07 A A 348 348 Isomer 1 Isomer 2 62

347.5 1.07 A 348 Isomer 1 63

415.7 2.51 A 416 64

385.5 1.39 A 386 65

329.5 6.62 L 330 66

343.5 6.97 L 344 67

343.5 7.15 L 344 68

343.5 7.17 L 344 69

343.5 7.11 L 344 70

345.5 3.25 C 346 71

347.5 5.60 L 348 72

351.5 8.26 L 353 Preparation (6S)-50D was used 73

357.5 7.83 L 358 74

357.5 7.65 L 358 75

357.5 7.73 L 358 76

359.5 6.50 L 360 77

359.6 3.33 C 360 78

361.5 5.97 L 362 79

365.5 7.64 L 366 Preparation (6S)-50D was used 80

365.5 8.78 L 366 81

365.5 7.76 L 366 Preparation (6S)-50D was used 82

365.5 8.84 L 366 83

365.5 8.75 L 366 84

369.5 5.09 L 370 85

371.5 6.89 L 372 86

371.5 6.80 L 372 87

373.5 6.97 L 374 88

379.5 7.51 L 380 89

381.5 2.88 C 382 90

381.5 3.01 C 382 91

382.5 4.23 L 383 92

382.5 4.22 L 383 Preparation (6S)-50D was used 93

385.5 2.96 C 386 94

393.6 8.49 L 394 95

393.6 8.58 L 394 96

395.5 6.98 L 396 97

395.5 7.11 L 396 Preparation (6S)-50D was used 98

395.5 7.03 L 396 Preparation (6S)-50D was used 99

395.5 6.97 L 396 100

395.5 7.07 L 396 101

395.5 7.56 L 396 102

399.5 6.50 L 360 103

399.5 8.21 L 400 104

399.5 7.12 L 400 105

399.5 7.56 L 400 106

399.5 7.04 L 400 107

399.6 7.72 L 400 108

399.6 7.82 L 400 109

408.6 5.50 L 409 110

411.5 2.86 C 412 111

413.6 2.34 A 414 112

413.6 2.15 A 414 113

416.0 8.77 L 416 114

416.0 7.63 L 416 115

416.0 7.25 L 416 116

415.9 8.02 L 416 117

417.5 3.11 C 418 118

422.6 5.88 L 523 119

423.5 7.14 L 424 120

435.5 8.26 L 436 121

436.6 6.21 L 437 122

437.6 3.64 C 438 123

439.5 7.89 L 440 124

464.7 6.93 L 465 125

464.7 6.99 L 465

The examples in Table 6 were prepared according to the generalprocedures of Examples 50 and 51.

TABLE 6 HPLC Ex. ret. time HPLC MS No. Structure MW (min.) condition(M⁺¹) Comment 126

333.5 0.28 A 334 Isomer 1 127

333.5 0.25 A 334 Isomer 2 128

361.5 0.44 A 362 Isomer 1 129

361.5 0.66 A 362 Isomer 2

The Examples in Table 7 were prepared according to the generalprocedures of Examples 30 and 31.

TABLE 7 HPLC Ex. ret. time HPLC MS No. Structure MW (min.) condition(M⁺¹) Comment 130 131

361.5 361.5 7.41 7.46 L L 362 362 Isomer 1 Isomer 2 132 133

359.2 359.2 6.18 6.20 L L 360 360 Isomer 1 Isomer 2 134 135

347.5 347.5 7.14 6.80 L L 348 348 Isomer 1 Isomer 2 136 137

357.5 357.5 8.07 8.97 L L 358 358 Isomer 1 Isomer 2

The Examples in Table 8 were prepared according to the generalprocedures of Examples 50 and 51.

TABLE 8 HPLC Ex. ret. time HPLC MS No. Structure MW (min.) condition(M⁺¹) Comment 138

367.5 1.72 A 368 Isomer 1 139

367.5 1.73 A 368 Isomer 2 140

347.6 1.53 A 348 Isomer 1 141

347.6 1.53 A 348 Isomer 2 142

333.5 3.15 C 334 143

333.5 3.12 C 334 144

347.6 3.30 C 348 145 146

347.6 347.6 8.91 8.90 L L 348.1 348.1 Isomer 1 Isomer 2 147 148

367.6 367.6 8.56 8.55 L L 368.1 368.1 Isomer 1 Isomer 2 149

347.6 3.26 C 348 150

361.6 2.05 A 362 151

361.6 3.43 C 362 152

361.6 3.43 C 362 153

361.6 3.49 C 362 154

361.6 3.47 C 362 155

361.6 3.40 C 362 156

368.5 2.42 C 369 157

368.5 1.66 C 369 158

375.6 3.59 C 376 159

375.6 3.49 C 376 160

381.6 3.40 C 382 161

382.6 2.13 C 283 162

391.7 2.03 A 392 163

395.6 3.38 C 396 164

395.6 3.36 C 396 165

395.6 3.63 C 396 166

396.6 0.99 A 397 167

396.6 1.79 C 397 168

397.6 1.36 A 398 169

397.6 2.56 C 398 170

397.6 3.05 C 398 171

397.6 3.21 C 398 172

397.6 3.21 C 398 173

409.6 3.58 C 410 174

425.7 2.16 A 426 175

427.6 3.03 C 428

Example 176((1R,3S)-1-amino-3-((S)-6-(2-(isobutylthio)ethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 176A:2-((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)ethyl4-methylbenzenesulfonate

(5R,7S)-7-((S)-6-(2-hydroxyethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(130 mg, 0.412 mmol) was dissolved in dry pyridine (1 mL) andp-toluenesulfonyl chloride (236 mg, 1.236 mmol) was added in oneportion. The resulting mixture was reacted at room temperature for 2 h.The solvent was removed in vacuo. The residue was dissolved in DCM andloaded onto column. Flash chromatography purification (0->100% ethylacetate in DCM) afforded2-((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)ethyl 4-methylbenzenesulfonate (169 mg, 0.360 mmol) as a solid. HPLCretention time=3.46 min (condition C); LC/MS M⁺¹=470.

Example 176

To a stirred mixture of isobutylmercaptan (0.021 mL, 0.192 mmol),2-((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)ethyl4-methylbenzenesulfonate (30 mg, 0.064 mmol), and dioxane (0.5 mL) wasadded 2N aqueous NaOH (0.096 mL, 0.192 mmol) at 0° C. under nitrogen.The resulting mixture was stirred at the same temperature for 15 min andat 60° C. for 6 h. Next, 2N aqueous NaOH (0.639 mL, 1.278 mmol) wasadded and the resulting mixture was stirred at 90° C. under nitrogenovernight. The mixture was extracted with ethyl acetate (4×1 mL) and thecombined ethyl acetate extracts were dried (Na₂SO₄) and concentrated.Purification using reverse phase HPLC (Phenomenex Luna 5p 30×100 mm(Axia); gradient over 8 min from 30 to 100% of solvent B; solvent A: 10%MeOH: 90% H₂O: 0.1% TFA; solvent B: 90% MeOH, 10% H₂O, 0.1% TFA),concentration, basification with 2N aqueous NaOH and extraction withethyl acetate gave((1R,3S)-1-amino-3-((S)-6-(2-(isobutylthio)ethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(22 mg, 0.057 mmol) as a white solid. HPLC retention time=3.39 min(condition C); LC/MS M+=362. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.10-6.87(m, 3H), 3.57-3.38 (m, 2H), 3.15-2.96 (m, 1H), 2.92-2.75 (m, 3H),2.68-2.58 (m, 2H), 2.48-2.37 (m, 3H), 2.29 (dd, J=13.1, 7.5 Hz, 1H),2.15-2.02 (m, 1H), 2.00-1.37 (m, 10H), 1.02 (d, J=6.6 Hz, 6H).

The Examples in Table 9 were prepared according to the general procedureof Example 176.

TABLE 9 HPLC Ex. ret. time HPLC MS No. Structure MW (min.) condition(M⁺¹) Comment 177

333.5 3.04 C 334 178

333.5 3.12 C 334 179

347.6 3.24 C 348 180

347.6 3.25 C 348 181

347.6 3.24 C 348 182

347.6 3.29 C 348 183

361.6 3.41 C 362

Example 184((1R,3S)-1-amino-3-((S)-6-(2-isobutoxyethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a stirred mixture of 2-methylpropan-1-ol (0.3 mL, 3.25 mmol) and2-((S)-6-((5R,7S)-2-oxo-3-oxa-1l-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)ethyl4-methylbenzenesulfonate (40 mg, 0.085 mmol) was added 1N THF solutionof potassium tert-butoxide (0.852 mL, 0.852 mmol) at 0° C. undernitrogen. The resulting mixture was at 70° C. for 6 hr before 2 Naqueous NaOH (0.426 mL, 0.852 mmol) was added. The mixture wasconcentrated to remove THE. Dioxane (0.5 mL) was added and the mixturewas stirred at 90° C. under nitrogen overnight. The mixture wasextracted with ethyl acetate (4×1 mL). The combined ethyl acetateextracts were dried (Na₂SO₄) and concentrated. Purification usingreverse phase HPLC (Phenomenex Luna 5u 30×100 mm (Axia); gradient over 8min from 30 to 10000 of solvent B; solvent A: 100% MeOH: 90% H₂O: 0.10%TFA; solvent B: 90% MeOH, 10% H₂O, 0.1% TFA), concentration,basification with 2N NaOH, and extraction with ethyl acetate gave((1R,3S)-1-amino-3-((S)-6-(2-isobutoxyethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(27 mg, 0.076 mmol) as a solid. HPLC retention time=3.41 min (conditionC); LC/MS M⁺¹=346. ¹H NMR (400 MHz, CTLOROFORM-d) δ 7.00-6.94 (m, 3H),3.52 (t, J=6.7 Hz, 2H), 3.45 (br, 2H), 3.18 (d, J=6.8 Hz, 2H), 3.08-2.96(m, 1H), 2.88-2.75 (m, 3H), 2.41 (dd, J=16.4, 10.5 Hz, 1H), 2.26 (dd,J=13.2, 7.9 Hz, 1H), 2.11-2.00 (m, 1H), 1.99-1.34 (m, 10H), 0.90 (d,J=6.6 Hz, 6H).

The Examples in Table 10 were prepared according to the generalprocedure of Example 184.

TABLE 10 HPLC Ex. ret. time HPLC MS No. Structure MW (min.) condition(M⁺¹) Comment 185 186

303.4 303.4 5.58 5.57 L L 304 304 187

317.5 6.28 L 318 188

331.5 7.11 L 332 189

343.5 2.94 C 344 190

343.5 2.83 C 344 191

345.5 3.22 C 346 192

345.5 3.41 C 346 193 194

395.5 395.5 7.50 7.50 L L 396 396 Isomer 1 Isomer 2

Example 195((1R,3S)-1-amino-3-((S)-6-(4-methoxy-2-methylbenzyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 195A:(5R,7S)-7-((S)-6-(4-methoxy-2-methylbenzyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (35 mg, 0.077 mmol) and copper(I) bromide (33.1mg, 0.230 mmol) in THF (3 mL) was added(4-methoxy-2-methylphenyl)magnesium bromide (4610 μl, 2.305 mmol) at−78° C. The reaction mixture was stirred at −78° C. and allowed to warmto room temperature over 16 h. The reaction mixture was diluted withsaturated NH₄Cl and water and extracted with EtOAc. The organic layerwas collected, dried over Na₂SO₄, concentrated on the rotavapor to giveto give(5R,7S)-7-((S)-6-(4-methoxy-2-methylbenzyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(20 mg) as a white solid. LC/MS M⁺¹=406.

Example 195

To a solution of(5R,7S)-7-((S)-6-(4-methoxy-2-methylbenzyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(20 mg, 0.049 mmol) in dioxane (3 mL) and water (1 mL) was added LiOH(11.81 mg, 0.493 mmol). The reaction mixture was stirred at 100° C. for16 h to give the crude product which was purified on a prep HPLC. HPLCconditions: Phenomenex Luna 5 micron C18 column (21.2×100 mm); MeOH(0.1% TFA)/water (0.1% TFA); 0%-100% gradient over 15 minutes; 20mL/min. Isolated fractions with correct mass were collected andfreeze-dried overnight. Recovered 10 mg of((1R,3S)-1-amino-3-((S)-6-(4-methoxy-2-methylbenzyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanolTFA. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.20-6.35 (m, 6H), 3.6 (s, 3H), 3.5(m, 2H), 3.2-2.6 (m, 5H), 2.4 (m, 1H), 2.3 (s, 3H), 2.2 (m, 1H), 2.2-1.5(m, 7H). LC/MS M⁺¹=380.

The Examples in Table 11 were prepared according to the generalprocedure of Example 195.

TABLE 11 HPLC ret. Ex. time HPLC MS No. Structure MW (min.) method (M⁺¹)Comment 195

379.5 7.52 L 380 196

379.5 7.74 L 380 Preparation (6S)-50D was used 197

379.5 8.80 L 380 Preparation (6S)-50D was used 198

379.5 8.81 L 380 199

379.5 7.13 L 380 200

379.5 8.81 L 380 201

379.5 7.12 L 380 202

395.5 6.64 L 380 203

383.5 7.28 L 384 204

327.5 8.33 L 328 205

341.5 8.80 L 342 206

371.6 7.30 L 372 207

357.5 6.73 L 358 208

393.6 8.07 L 394 209

393.6 8.01 L 394 210

353.3 5.56 L 354 211

358.6 4.28 L 359 212

353.5 5.27 L 354 213

420.6 6.12 L 421 214

393.6 8.39 L 394 215

393.6 8.19 L 394 216

505.4 8.87 L 506 217 218

353.5 353.5 3.77 3.77 L L 354 354 219

365.5 7.45 L 366 220

365.5 7.26 L 366 221

365.5 7.25 L 366 222

365.5 7.31 L 366 223

365.5 7.24 L 366 224

336.5 3.19 L 337 Isomer 1 225

336.5 3.20 L 337 Isomer 2

Examples 226 and 227((1R,3S)-1-amino-3-(2-hexyl-2,3-dihydro-1H-inden-5-yl)cyclopentyl)methanol

Preparation 226A:(5R,7S)-7-(6-phenyl-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-3,4-dihydronaphthalen-2-yltrifluoromethanesulfonate (75 mg, 0.180 mmol), triphenylphosphine (10mg, 0.038 mmol), and acetylacetone cobalt (III) salt (4 mg, 0.011 mmol)in THF (5 mL) was added phenylmagnesium bromide (0.539 mL, 0.539 mmol).The reaction mixture was stirred for 3 hours and during this time wasallowed to warm to room temperature. The reaction was quenched withwater and the mixture was diluted with ethyl acetate and washed withsaturated NaCl. The organic layer was dried with MgSO₄, filtered andconcentrated. The crude material was purified on a silica gel cartridge(24 g) using an EtOAc/Hex gradient (0-100% EtOAc over 13 CV) to afford55 mg of(5R,7S)-7-(6-phenyl-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.HPLC retention time=1.06 min (condition G); LC/MS M⁺¹=346.

Preparation 226B:(5R,7S)-7-(6-phenyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of(5R,7S)-7-(6-phenyl-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(55 mg, 0.159 mmol) in MeOH (10 mL) was added Pearlman's Catalyst (22.36mg, 0.159 mmol). The mixture was hydrogenated under a balloon of H₂ forone hour. The mixture was filtered to remove the catalyst andconcentrated to afford 38 mg of(5R,7S)-7-(6-phenyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.The individual isomers were separated using a CHIRALPAK® AD-H columnunder SFC conditions (50% MeOH in CO₂). Isomer 1 (226B, 12 mg),retention time on chiral HPLC, 10.3 min; MS (m+1)=348. Isomer 2 (227B,12 mg), retention time on chiral HPLC, 13.3 min.; MS (m+1)=348. Theabsolute stereochemistry of the isomers was not determined.

Examples 226 and 227

To a mixture of(5R,7S)-7-(6-phenyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(12 mg, 0.035 mmol, 226B, isomer 1) in dioxane (2 mL) was added 2N NaOH.The reaction mixture was heated at 100° C. overnight, cooled, and thenacidified with TFA. The solvents were removed, MeOH (1.8 mL) was added,and the mixture was filtered to remove solids and purified by HPLC. HPLCconditions: Phenomenex Luna 5 micron C18 column (30×100 mm); MeCN (0.1%TFA)/water (0.1% TFA); 20%-100% gradient over 15 minutes; 30 mL/min.Recovered 8 mg of((1R,3S)-1-amino-3-(6-phenyl-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(Isomer 1, Example 226). HPLC retention time=8.22 min (condition H);LC/MS M⁺¹=322; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.40-7.25 (m, 4H),7.25-7.15 (m, 1H), 7.11-6.97 (m, 3H), 3.81-3.55 (m, 2H), 3.24-3.06 (m,1H), 3.03-2.79 (m, 5H), 2.45 (ddd, J=13.4, 7.1, 1.1 Hz, 1H), 2.23-2.06(m, 2H), 2.04-1.86 (m, 4H), 1.75 (t, J=12.7 Hz, 1H); MS (m+1)=322.

To a mixture of(5R,7S)-7-(6-phenyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(12 mg, 0.035 mmol, 227B, isomer 2) in dioxane (2 mL) was added 2N NaOH.The reaction mixture was heated at 100° C. overnight, cooled, and thenacidified with TFA. The solvents were removed, MeOH (1.8 mL) was added,and the mixture was filtered to remove solids and purified by HPLC. HPLCconditions: Phenomenex Luna 5 micron C18 column (30×100 mm); MeCN (0.1%TFA)/water (0.1% TFA); 20%-100% gradient over 15 minutes; 30 mL/min.Recovered 4.5 mg of((1R,3S)-1-amino-3-(6-phenyl-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(Example 227, Isomer 2). HPLC retention time=8.24 min (condition H);LC/MS M⁺¹=322; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.36-7.27 (m, 4H),7.24-7.17 (m, 1H), 7.05 (s, 3H), 3.73-3.58 (m, 2H), 3.21-3.08 (m, 1H),3.05-2.82 (m, 5H), 2.44 (ddd, J=13.4, 7.1, 1.1 Hz, 1H), 2.21-2.07 (m,2H), 2.04-1.88 (m, 4H), 1.75 (t, J=12.7 Hz, 1H); MS (m+1)=322.

The Examples in Table 12 were prepared according to the generalprocedure of Examples 226 and 227.

TABLE 12 HPLC Ex. ret. time HPLC MS No. Structure MW (min.) condition(M⁺¹) Comment 228 229

335.5 335.5 8.87 8.92 H H 336 336 Isomer 1 Isomer 2 230 231

349.5 349.5 9.15 9.16 H H 350 350 Isomer 1 Isomer 2 232 233

322.5 322.5 3.78 0.5  H G 323 323 Isomer 1 Isomer 2 234 235

315.5 315.5 9.57 9.67 L L 316 316 Isomer 1 Isomer 2 236 237

301.5 301.5 9.05 9.05 L L 302 302 Isomer 1 Isomer 2

Examples 238 and 239((1R,3S)-3-(6-(2-(allyloxy)ethoxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-1-aminocyclopentyl)methanol

Preparation 238A:(5R,7S)-7-(3′,4′-dihydro-1′H-spiro[[1,3]dioxolane-2,2′-naphthalen]-6′-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To the mixture of(5R,7S)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(200 mg, 0.701 mmol) and ethane-1,2-diol (870 mg, 14.02 mmol) in MeCN (5ml), p-toluenesulfonic acid (26.7 mg, 0.140 mmol) was added and themixture was stirred at room temperature for 16 h. The mixture wasdiluted with EtOAc (50 ml), the organic layer was washed with saturatedNaHCO₃(3×20 ml), dried with Na₂SO₄ and concentrated under reducedpressure to give 205 mg of crude(5R,7S)-7-(3′,4′-dihydro-1′H-spiro[[1,3]dioxolane-2,2′-naphthalen]-6′-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.HPLC retention time=2.58 min (condition B); LC-MS M⁺¹=330.

Preparation 238B:(5R,7S)-7-(6-(2-hydroxyethoxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To the mixture of(5R,7S)-7-(3′,4′-dihydro-1′H-spiro[[1,3]dioxolane-2,2′-naphthalen]-6′-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(50 mg, 0.152 mmol) and BF₃.OEt₂ (192 μl, 1.52 mmol) in THF (5 ml),NaCNBH₄ (76 mg, 1.21 mmol) was added. The mixture was stirred at roomtemperature overnight. The reaction was quenched with water (1 ml) at 0°C. The mixture was diluted with EtOAc (40 ml), washed with saturatedNaHCO₃(2×20 ml), dried with Na₂SO₄ and concentrated under reducedpressure to give 50 mg of(5R,7S)-7-(6-(2-hydroxyethoxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.HPLC retention time=2.42 min (condition B); LC-MS M⁺¹=332.

Preparation 238C:2-((6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)oxy)ethyl4-methylbenzenesulfonate

To the mixture of(5R,7S)-7-(6-(2-hydroxyethoxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(300 mg, 0.905 mmol) in dry pyridine (5 ml), 4-methylbenzene-1-sulfonylchloride (518 mg, 2.72 mmol) was added in one portion at 0° C. Theresulting mixture was stirred at room temperature for 1 h, the mixturewas diluted with EtOAc (80 ml), washed with saturated NaHCO₃(3×30 ml),dried with Na₂SO₄ and concentrated under reduced pressure. The residuewas purified with silica gel cartridge (40 g) using an EtOAc/Hexgradient (0-65% EtOAc over 40 minutes) to provide 360 mg2-((6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)oxy)ethyl4-methylbenzenesulfonate. HPLC retention time=3.33 min (condition B);LC-MS M⁺¹=486. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.77 (d, J=8.1 Hz, 2H),7.40 (d, J=7.9 Hz, 2H), 7.04-6.94 (m, 3H), 4.42-4.26 (m, 2H), 4.17 (t,J=4.5 Hz, 2H), 3.73 (dt, J=9.5, 4.5 Hz, 3H), 3.08-2.81 (m, 3H),2.74-2.60 (m, 2H), 2.45 (s, 3H), 2.34-2.27 (m, 1H), 2.09 (s, 2H), 1.95(s, 3H), 1.82-1.75 (m, 2H).

Preparations 238D and 239D:(5R,7S)-7-(6-(2-(allyloxy)ethoxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To the mixture of2-((6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)oxy)ethyl4-methylbenzenesulfonate (80 mg, 0.165 mmol) and prop-2-en-1-ol (28.7mg, 0.494 mmol) in 2 ml of THF, KOtBu (92 mg, 0.824 mmol) was added andthe mixture was stirred at room temperature for 16 h, then at 65° C. for1.5 h. The mixture was quenched with water (1 ml) at 0° C., diluted withEtOAc (40 ml), washed with saturated NaHCO₃(2×20 ml), dried with Na₂SO₄and concentrated under reduced pressure to give 60 mg(5R,7S)-7-(6-(2-(allyloxy)ethoxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.The individual isomers were separated using a Chiral OD-H 25×3 cm ID,Sum under SFC conditions (20% MeOH in CO₂).

Preparation 238D: Isomer 1 (10 mg), HPLC retention time=3.20 min(condition B); MS (m+1)=372; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.05-6.94(m, 3H), 6.00-5.82 (m, 1H), 5.33-5.08 (m, 2H), 4.43-4.26 (m, 2H), 4.03(dt, J=5.6, 1.4 Hz, 2H), 3.89-3.58 (m, 5H), 3.12-2.85 (m, 3H), 2.83-2.69(m, 2H), 2.29 (dd, J=13.0, 7.0 Hz, 1H), 2.18-2.02 (m, 3H), 2.01-1.74 (m,4H).Preparation 239D: Isomer 2 (8 mg), HPLC retention time=3.19min(condition B); MS (m+1)=372. ¹H NMR (400 MHz, METHANOL-d₄) δ7.11-6.94 (m, 3H), 6.01-5.84 (m, 1H), 5.35-5.11 (m, 2H), 4.47-4.25 (m,2H), 4.03 (dt, J=5.6, 1.4 Hz, 2H), 3.88-3.56 (m, 5H), 3.13-2.87 (m, 3H),2.83-2.67 (m, 2H), 2.29 (dd, J=13.0, 7.0 Hz, 1H), 2.17-1.71 (m, 7H). Theabsolute stereochemistry of the isomers was not determined.

Examples 238 and 239

To the mixture of(5R,7S)-7-(6-(2-(allyloxy)ethoxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(Preparation 238D, 10 mg, 0.027 mmol) and LiOH—H₂O (13.6 mg, 15 eq) indioxane (1.5 ml) and water (0.5 ml) was heated at 100° C. for 16 h.After cooling, the mixture was diluted with DCM (50 ml) and water (20ml), the organic layer was separated and the aqueous layer was addedsaturated NaHCO₃ (10 ml) and extracted with DCM (30 ml). The combinedDCM mixture was dried with Na₂SO₄, concentrated under vacuo and purifiedwith preparative HPLC: column Phenomenex Luna C18 5u 21.2×100 mm.Solvent A: 10% MeOH-90% H₂O-0.1% TFA; Solvent B: 90% MeOH-10% H₂O-0.1%TFA. Gradient time=15 min. Start B=0%, Final B 100%. Stop time 25 min.The desired peak was collected, basified to approximately pH 8 withsaturated NaHCO₃, the solvent was removed under reduced pressure and theaqueous layer was extracted with DCM (3×30 ml), which was dried withNa₂SO₄, concentrated under reduced pressure, redissolved in MeCN (2 ml)and water (1 ml) and lyophilized for overnight to give Example 238 (6 mgof isomer 1)((1R,3S)-3-(6-(2-(allyloxy)ethoxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-1-aminocyclopentyl)methanol. HPLC retention time=8.0 min (condition L); LC-MS M⁺¹=346; ¹HNMR (400 MHz, METHANOL-d₄) δ 7.08-6.94 (m, 3H), 5.92 (ddt, J=17.2, 10.7,5.4 Hz, 1H), 5.34-5.12 (m, 2H), 4.03 (dt, J=5.6, 1.5 Hz, 2H), 3.89-3.58(m, 7H), 3.21-2.86 (m, 3H), 2.82-2.71 (m, 2H), 2.48-2.36 (m, 1H),2.19-2.02 (m, 2H), 2.01-1.81 (m, 4H), 1.72 (t, J=12.7 Hz, 1H).

Example 239 (5 mg of isomer 2)((1R,3S)-3-(6-(2-(allyloxy)ethoxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-1-aminocyclopentyl)methanolwas prepared similarly from 8 mg of Preparation 239D of(5R,7S)-7-(6-(2-(allyloxy)ethoxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.HPLC retention time=7.99 min (condition L); LC-MS M⁺¹=346; ¹H NMR (400MHz, METHANOL-d₄) δ 7.08-6.95 (m, 3H), 6.02-5.82 (m, 1H), 5.33-5.12 (m,2H), 4.03 (dt, J=5.5, 1.4 Hz, 2H), 3.88-3.54 (m, 7H), 3.14-2.87 (m, 3H),2.82-2.66 (m, 2H), 2.33 (dd, J=13.2, 6.4 Hz, 1H), 2.15-2.02 (m, 2H),1.98-1.80 (m, 4H), 1.65 (t, J=12.5 Hz, 1H). The Examples in Table 13were prepared according to the general procedure of Examples 238 and239.

TABLE 13 HPLC ret. HPLC Ex. time me- MS No. Structure MW (min.) thod(M⁺¹) Comment 240 241

373.5 373.5 8.71 8.73 L L 374 374 Isomer 1 Isomer 2 242

395.5 8.94 L 396 Isomer 1 243

395.5 8.95 L 396 Isomer 2 244

395.5 8.94 L 396 Isomer 1 245

395.5 8.95 L 396 Isomer 2 246

359.5 7.06 L 360 OH-Is#2 247

387.6 8.03 L 388 248

387.6 6.87 L 388 249

359.5 6.05 L 360 250

375.6 7.00 L 376 251

375.6 6.97 L 376 252

361.5 6.51 L 362 253

389.6 7.50 L 390 254

402.6 5.08 L 403 255

401.6 6.33 L 402 256

359.5 5.88 L 360 257

373.5 7.81 L 374 258

373.5 6.58 L 374 259

357.4 5.75 L 358 260

401.5 6.37 L 402 261

357.4 5.76 L 358 262

402.5 5.09 L 403 263

363.5 6.78 L 364 264

401.5 6.29 L 402 265

395.5 7.10 L 396 266 267

361.5 361.5 8.73 8.74 L L 362 362 Isomer 1 Isomer 2

Examples 268 and 269((1R,3S)-1-amino-3-(6-(4-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a mixture of6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-3,4-dihydronaphthalen-2-yltrifluoromethanesulfonate (100 mg, 0.240 mmol), copper(I) iodide (4.56mg, 0.024 mmol), and bis(triphenylphosphine)palladium(II) chloride(16.82 mg, 0.024 mmol) in TEA (3 mL) was added1-ethynyl-4-methoxybenzene (63.3 mg, 0.479 mmol). The reaction mixturewas heated at 60° C. for 1 hour. The reaction mixture was diluted withethyl acetate and washed with 1M HCl. The organic layer was dried withMgSO₄, filtered and concentrated. The crude material was purified on asilica gel cartridge (24 g) using an EtOAc/Hex gradient (0-100% EtOAcover 13 CV) to afford(5R,7S)-7-(6-((4-methoxyphenyl)ethynyl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(68 mg, 0.170 mmol). LC/MS M+1=402.

To a mixture of(5R,7S)-7-(6-((4-methoxyphenyl)ethynyl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(62 mg, 0.155 mmol) in MeOH (5 mL) was added Pearlman's Catalyst (10.90mg, 0.078 mmol). The flask was charged with hydrogen and hydrogenatedunder a balloon for 2 hours. The catalyst was filtered away and themixture was concentrated in vacuo to afford 45 mg of(5R,7S)-7-(6-(4-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.Two diastereomers were separated under SFC conditions on a Chiral AS-H25×3 cm ID, 5 μm column and eluting with 60/40 CO₂/MeOH at 85.0 mL/min.Each isomer was taken to the next step. LC/MS M⁺¹=406.

Example 268: To a mixture of(5R,7S)-7-(6-(4-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(18 mg, 0.044 mmol) in DMSO (1 mL) and MeOH (1 mL) was added 1N NaOH(0.5 mL). The reaction mixture was heated at 90° C. overnight. Themixture was acidified with TFA followed by the removal of most solvent.The mixture was filtered and purified by HPLC. HPLC conditions:Phenomenex Luna 5 micron C18 column (30×100 mm); MeCN (0.1% TFA)/water(0.1% TFA); 20%-100% gradient over 15 minutes; 30 mL/min. Fractions withcorrect mass were isolated and freeze-dried overnight to afford((1R,3S)-1-amino-3-(6-(4-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (14 mg, 0.026 mmol). ¹H NMR in CD₃OD (400 MHz, METHANOL-d₄) δ 7.13(d, J=8.6 Hz, 2H), 7.04-6.96 (m, 3H), 6.84 (d, J=8.8 Hz, 2H), 3.77 (s,3H), 3.71-3.55 (m, 2H), 3.19-3.01 (m, 1H), 2.95-2.73 (m, 3H), 2.73-2.63(m, 2H), 2.42 (dd, J=14.2, 7.8 Hz, 2H), 2.21-2.06 (m, 1H), 2.05-1.85 (m,4H), 1.80-1.59 (m, 4H), 1.43 (dtd, J=12.8, 10.5, 6.1 Hz, 1H). MS(m+1)=380. HPLC Peak RT=10.16 min. (Condition L). Purity=92%.

Example 269

To a mixture of(5R,7S)-7-(6-(4-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(17 mg, 0.044 mmol) in DMSO (1 mL) and MeOH (1 mL) was added 1N NaOH(0.5 mL). The mixture was heated at 90° C. overnight. The mixture wasacidified with TFA followed by the removal of most solvent. The mixturewas filtered and purified by HPLC. HPLC conditions: Phenomenex Luna 5micron C18 column (30×100 mm); MeCN (0.1% TFA)/water (0.1% TFA);20%-100% gradient over 15 minutes; 30 mL/min. Fractions with the correctmass were isolated and freeze-dried overnight to afford((1R,3S)-1-amino-3-(6-(4-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (14 mg, 0.026 mmol). ¹H NMR in CD₃OD (400 MHz, METHANOL-d₄) δ 7.13(d, J=8.6 Hz, 2H), 7.04-6.96 (m, 3H), 6.84 (d, J=8.8 Hz, 2H), 3.77 (s,3H), 3.70-3.55 (m, 2H), 3.19-3.03 (m, 1H), 2.97-2.74 (m, 3H), 2.73-2.62(m, 2H), 2.42 (dd, J=14.1, 7.7 Hz, 2H), 2.21-2.06 (m, 1H), 2.05-1.87 (m,4H), 1.81-1.58 (m, 4H), 1.43 (dtd, J=12.7, 10.6, 5.9 Hz, 1H). MS(m+1)=380. HPLC Peak RT=10.16 min. (Condition L) Purity=99%.

The Examples in Table 14 were prepared according to the generalprocedure of Examples 268 and 269.

TABLE 14 HPLC ret. HPLC Ex. Time condi- MS No. Structure MW (min.) tion(M⁺¹) Comment 270

317.5 7.18 L 318 Isomer 1 271

317.5 7.16 L 318 Isomer 2 272

331.5 7.64 L 332 Isomer 1 273

331.5 7.64 L 332 Isomer 2 274

359.6 7.58 L 360 Isomer 1 275

359.6 7.58 L 360 Isomer 2 276 277

345.5 345.5 8.18 8.17 L L 346.4 346.4 Isomer 1 Isomer 2

Example 278((1R,3S)-1-amino-3-((S)-6-(3-isopropoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 278A:(5R,7S)-7-((R)-6-(but-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

A diethyl ether solution (1M) of allylmagnesium bromide (4.39 mL, 4.39mmol) was added to a stirred mixture of copper(I) bromide (63.0 mg,0.439 mmol),((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (100 mg, 0.220 mmol) and anhydroustetrahydrofuran (2 mL) at −78° C. under nitrogen. The mixture wasstirred at −78° C. for 20 min before the temperature was slowly raisedto room temperature. The mixture was stirred at room temperature for 16hr. Saturated aqueous NH₄Cl solution (3 mL) was added slowly to quenchthe reaction. Ethyl acetate (4 mL) and water (1 mL) were added. Theaqueous layer was separated and extracted with ethyl acetate (2×3 mL).The combined organic solutions were dried over sodium sulfate andconcentrated under reduced pressure to give(5R,7S)-7-((R)-6-(but-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(90 mg, 0.277 mmol). LC/MS M⁺¹=326.

Preparation 278B:3-((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propanal

To a clear solution of(5R,7S)-7-((R)-6-(but-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(0.09 g, 0.277 mmol) in THF (1.5 mL) were sequentially added 50% NMO inwater (0.115 mL, 0.553 mmol) and 4% osmium tetroxide in water (0.051 mL,8.30 μmol) at room temperature. The solution was vigorously stirred atroom temperature overnight. Additional 50% NMO in water (0.06 mL) wasadded. The solution was vigorously stirred at room temperature for 1day. Sodium periodate (0.237 g, 1.106 mmol) in H₂O (1 mL) was added andthe mixture was stirred vigorously at room temperature under nitrogenfor 30 min. The mixture was extracted with ethyl acetate (3×2 mL). Thecombined ethyl acetate extracts were dried (Na₂SO₄) and concentrated.Flash chromatography purification (4 g silica gel column, gradientelution from 15 to 100% of ethyl acetate in hexanes) afforded3-((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propanal(63 mg, 0.192 mmol) as a solid. HPLC retention time=2.92 min (conditionC); LC/MS M⁺¹=328.

Preparation 278C:(5R,7S)-7-((S)-6-(3-isopropoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a stirred solution of3-((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propanal(21 mg, 0.064 mmol), isopropoxytrimethylsilane (0.057 mL, 0.321 mmol),and triethylsilane (0.051 mL, 0.321 mmol) in nitromethane (1 mL) wasadded ferric chloride (1.040 mg, 6.41 μmol) at 0° C. under nitrogen. Themixture was stirred at 0° C. for 15 min and at room temperature for 30min before being concentrated. The residue was mixed with saturatedaqueous sodium bicarbonate solution (1 mL) and extracted with ethylacetate (3×1 mL). The combined ethyl acetate extracts were dried(Na₂SO₄) and concentrated under reduced pressure to give(5R,7S)-7-((S)-6-(3-isopropoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(23 mg, 0.062 mmol) as a solid. LC/MS M⁺¹=372.

Example 278

A mixture of(5R,7S)-7-((S)-6-(3-isopropoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(23 mg, 0.062 mmol), 2N aqueous NaOH (0.619 mL, 1.238 mmol), and dioxane(0.5 mL) was stirred at 90° C. under nitrogen overnight. The mixture wascooled and extracted with ethyl acetate (4×1 mL). The combined organicsolutions were dried over sodium sulfate and concentrated under reducedpressure. Purification using reverse phase HPLC (Phenomenex Luna 5p30×100 mm (Axia); gradient over 8 min from 30 to 100% of solvent B;solvent A: 10% MeOH: 90% H₂O: 0.1% TFA; solvent B: 90% MeOH, 10% H₂O,0.1% TFA), concentration, basification with 2N aqueous NaOH, andextraction with ethyl acetate gave((1R,3S)-1-amino-3-((S)-6-(3-isopropoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(21 mg, 0.052 mmol) as a white solid. HPLC retention time=3.04 min(condition C); LC/MS M⁺¹=346. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.03-6.93(m, 3H), 3.55 (dt, J=12.2, 6.1 Hz, 1H), 3.51-3.37 (m, 4H), 3.08-2.93 (m,1H), 2.89-2.71 (m, 3H), 2.38 (dd, J=16.2, 10.7 Hz, 1H), 2.26 (dd,J=13.0, 7.7 Hz, 1H), 2.04 (br. s., 1H), 1.98-1.60 (m, 7H), 1.56-1.45 (m,1H), 1.45-1.32 (m, 3H), 1.16 (d, J=6.2 Hz, 6H).

Example 279((1R,3S)-1-amino-3-((R)-6-(3-(oxetan-3-yloxy)propyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 279A:(5R,7S)-7-((R)-6-(3-hydroxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a stirred solution of3-((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propanal(162 mg, 0.495 mmol) (Example 278 step B) in 100% ethanol (8 mL) anddichloromethane (2 mL) was added NaBH₄ (18.72 mg, 0.495 mmol) at roomtemperature under nitrogen. The mixture was stirred at room temperaturefor 1 h. The mixture was concentrated. The reaction was quenched withsaturated aqueous NH₄Cl solution (1 mL) and water (1 mL) and the mixturewas extracted with ethyl acetate (4 mL, 2×1 mL). The combined organicsolutions were dried over sodium sulfate and concentrated under reducedpressure to give(5R,7S)-7-((R)-6-(3-hydroxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one (170 mg, 0.516 mmol) as a white solid. HPLC retentiontime=3.03 min (condition C); LC/MS M⁺¹=330.

Preparation 279B:3-((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propyl4-methylbenzenesulfonate

The above intermediate was prepared using the same procedure asPreparation 176A

Example 279

To a stirred mixture of3-((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propyl4-methylbenzenesulfonate (30 mg, 0.062 mmol) and oxetan-3-ol (0.06 mL,1.016 mmol) was added 1N THF solution of potassium tert-butoxide (0.620mL, 0.620 mmol) at 0° C. under nitrogen. The resulting mixture was atroom temperature for 5 h and 60° C. for 1 hr before 2 N aqueous NaOH(0.310 mL, 0.620 mmol) was added. The mixture was concentrated to removeTHF. Dioxane (0.5 mL) was added and the mixture was stirred at 70° C.under nitrogen for 15 hr and at 100° C. for 5 hr. The mixture was cooledand extracted with ethyl acetate (4×1 mL). The combined ethyl acetateextracts were dried (Na₂SO₄) and concentrated. Purification usingreverse phase HPLC (Phenomenex Luna 5p 30×100 mm (Axia); gradient over 9min from 20 to 100% of solvent B; solvent A: 10% MeOH: 90% H₂O: 0.1%TFA; solvent B: 90% MeOH, 10% H₂O, 0.1% TFA), concentration,basification with 2N NaOH, and extraction with ethyl acetate gave((1R,3S)-1-amino-3-((R)-6-(3-(oxetan-3-yloxy)propyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(19 mg, 0.045 mmol) as a solid. HPLC retention time=2.78 min (conditionC); LC/MS M⁺¹=360. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.05-6.87 (m, 3H),4.81-4.70 (m, 2H), 4.61 (t, J=6.2 Hz, 2H), 4.57-4.48 (m, 1H), 3.37 (t,J=6.6 Hz, 2H), 3.02 (br. s., 1H), 2.90-2.72 (m, 3H), 2.45-2.18 (m, 2H),2.13-1.61 (m, 10H), 1.56-1.31 (m, 4H).

The Examples in Table 15 were prepared according to the generalprocedure of Examples 278 and 279.

TABLE 15 HPLC ret. HPLC Ex. time condi- MS No. Structure MW (min.) tion(M⁺¹) Comment 280 281

343.5 343.5 6.96 6.93 L L 344 344 Isomer 1 Isomer 2 282

345.5 3.08 C 346 283

359.5 2.80 C 360 284 285

363.6 363.6 7.03 7.01 L L 364 364 Isomer 1 Isomer 2

Example 286((1R,3S)-1-amino-3-((S)-6-(2-(pyridin-2-yl)ethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 286A:6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carbaldehyde

Preparations 286B and 286C:(5R,7S)-7-((S)-6-ethynyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-oneand(5R,7S)-7-((R)-6-ethynyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carbaldehyde(718 mg, 2.4 mmol) and potassium carbonate (995 mg, 7.20 mmol) in MeOH(3 mL) was added dimethyl (1-diazo-2-oxopropyl) phosphonate (0.540 mL,3.60 mmol). The reaction mixture was stirred at room temperature for onehour. The reaction mixture was diluted with ethyl acetate and washedwith saturated NaCl. The organic layer was dried with MgSO₄, filteredand concentrated. The crude material was purified on a silica gelcartridge (40 g) using an EtOAc/Hex gradient (20-100% EtOAc over 12 CV)to afford 580 mg of(5R,7S)-7-6-ethynyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.The diastereomeric mixture was separated by SFC using a Chiralpak IC,25×3 cm ID, 5 μm column and eluting with 90/10 CO₂/MeOH at 85.0 mL/min.Peak 1 was isolated to afford(5R,7S)-7-((S)-6-ethynyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-azaspiro[4.4]nonan-2-one(225 mg, 0.762 mmol). Peak 2 was isolated to afford(5R,7S)-7-((R)-6-ethynyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(245 mg, 0.829 mmol). The absolute stereochemistry was determined byconverting Preparation 286B to Preparation 677B. Chiral HPLC analysisindicates compounds were identical and 286B was assigned as the Sstereochemistry at the alkynyl center. Preparation 286C was thenassigned the R configuration.

Example 286

An oven dried round bottom flask was charged with cesium carbonate (66.2mg, 0.203 mmol) and bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II) (3 mg, 4.24 μmol) under nitrogen. The mixture wasdegassed three times under vacuum, followed by the stepwise addition of2-bromopyridine (10 μl, 0.105 mmol),(5R,7S)-7-((R)-6-ethynyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(20 mg, 0.068 mmol), and acetonitrile (1 mL). The reaction mixture washeated at 80° C. overnight. Solvent was removed in vacuo and the residuewas dissolved in MeOH (2 mL). Pearlman's Catalyst (5 mg, 0.036 mmol) wasadded and the mixture was hydrogenated under a balloon of H₂ for 1 hour.The catalyst was removed by filtration. Next, 1N NaOH (2 mL) was addedto the filtrate and the mixture was heated at 95° C. for 6 hours. Themixture was acidified with TFA then filtered and purified by HPLC. HPLCconditions: Phenomenex Luna 5 micron C18 column (30×100 mm); MeCN (0.1%TFA)/water (0.1% TFA); 20%-100% gradient over 15 minutes; 30 mL/min.Fractions were isolated with the correct mass and freeze-dried overnightto afford((1R,3S)-1-amino-3-((S)-6-(2-(pyridin-2-yl)ethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(13 mg, 0.033 mmol). HPLC Peak RT=3.66 minutes (Condition L) purity=90%.MS (m+1)=351. ¹H NMR (400 MHz, METHANOL-d₄) δ 8.44 (dd, J=5.1, 0.9 Hz,1H), 7.77 (td, J=7.7, 1.8 Hz, 1H), 7.37 (d, J=7.9 Hz, 1H), 7.26 (ddd,J=7.5, 5.1, 1.1 Hz, 1H), 6.99 (s, 3H), 3.62-3.46 (m, 2H), 3.14-2.98 (m,1H), 2.93 (t, J=7.8 Hz, 2H), 2.89-2.69 (m, 2H), 2.45 (dd, J=16.3, 9.7Hz, 1H), 2.31 (dd, J=13.2, 6.4 Hz, 1H), 2.12-1.98 (m, 2H), 1.97-1.87 (m,3H), 1.87-1.72 (m, 4H), 1.63 (t, J=12.5 Hz, 1H), 1.46 (dtd, J=12.8,10.4, 5.9 Hz, 1H).

The Examples in Table 16 were prepared according to the generalprocedure of Example 286.

TABLE 16 HPLC ret. Ex. Time HPLC MS No. Structure MW (min.) condition(M⁺¹) 287

350.5 3.62 L 351 288

380.5 4.09 L 381 289

400.6 4.37 L 401 290

400.6 4.45 L 401 291

350.5 3.82 L 351 292

350.1 3.86 L 351 293

350.1 3.77 L 351 294

351.5 4.94 L 352 295

351.5 4.95 L 352 296

351.5 5.17 L 352 297

380.5 5.13 L 381 298

393.6 8.41 L 394 299

397.5 7.97 L 398 300

397.5 7.89 L 398 301

397.5 9.02 L 398 302

393.6 8.17 L 394 303

393.6 8.14 L 394 304

393.6 8.26 L 394

Example 305((1R,3S)-1-amino-3-((S)-6-(2-(pyridin-2-yl)ethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 305A:(5R,7S)-7-((R)-6-(((1-phenyl-1H-tetrazol-5-yl)sulfonyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (1 g, 2.195 mmol) and potassium carbonate(0.910 g, 6.59 mmol) in DMF (10 mL) was added1-phenyl-1H-tetrazole-5-thiol (0.782 g, 4.39 mmol). The reaction mixturewas heated at 80° C. overnight. The reaction mixture was diluted withethyl acetate and washed with saturated NaCl. The organic layer wasdried with MgSO₄, filtered and concentrated. The crude material waspurified on a silica gel cartridge (40 g) using an EtOAc/Hex gradient(0-100% EtOAc over 13 CV) to afford(5R,7S)-7-((R)-6-(((1-phenyl-1H-tetrazol-5-yl)thio)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(0.94 g, 2.036 mmol). LC/MS M⁺¹=462.

To hydrogen peroxide (8.32 mL, 81 mmol) at 0° C. was added ammoniummolybdate tetrahydrate (0.503 g, 0.407 mmol). The resulting solution wasadded to a mixture of(5R,7S)-7-((R)-6-(((1-phenyl-1H-tetrazol-5-yl)thio)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(0.94 g, 2.036 mmol) in THF (30 mL) at 0° C. The reaction mixture wasstirred overnight at room temperature.

The reaction mixture was diluted with ethyl acetate and washed withsaturated NaCl. The organic layer was dried with MgSO₄, filtered andconcentrated to afford(5R,7S)-7-((R)-6-(((1-phenyl-1H-tetrazol-5-yl)sulfonyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-oneone (1 g, 2.026 mmol) which was used without further purification. LC/MSM⁺¹=494.

Preparation 305B:(5R,7S)-7-((R)-6-((E)-2-fluoro-5-methoxystyryl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of 2-fluoro-5-methoxybenzaldehyde (30.0 mg, 0.194 mmol) and(5R,7S)-7-((R)-6-(((1-phenyl-1H-tetrazol-5-yl)sulfonyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(32 mg, 0.065 mmol) in THF was added KHMDS (0.259 mL, 0.259 mmol). Afterstirring at room temperature for 1 hour, the reaction was quenched withMeOH. The reaction mixture was purified by HPLC. HPLC conditions:Phenomenex Luna 5 micron C18 column (30×100 mm); MeCN (0.1% TFA)/water(0.1% TFA); 20%-100% gradient over 15 minutes; 30 mL/min. Fractions withcorrect mass were combined and freeze-dried overnight. Recovered(5R,7S)-7-((R)-6-((E)-2-fluoro-5-methoxystyryl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(6 mg, 0.014 mmol). LC/MS M⁺¹=422.

Example 305

To a mixture of(5R,7S)-7-((R)-6-((E)-2-fluoro-5-methoxystyryl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(6 mg, 0.014 mmol) in MeOH (2 mL) was added Pearlman's Catalyst (0.5 mg,3.56 μmol). The mixture was hydrogenated under a balloon of hydrogen for1 hour. The catalyst was removed by filtration. Next, 1 N NaOH (2 mL)was added and the mixture was heated to reflux overnight. The mixturewas cooled and acidified with TFA then purified by HPLC. HPLCconditions: Phenomenex Luna 5 micron C18 column (30×100 mm); MeCN (0.1%TFA)/water (0.1% TFA); 20%-100% gradient over 15 minutes; 30 mL/min.Fractions were isolated with the correct mass and freeze-dried overnightto afford(1R,3S)-1-amino-3-((S)-6-(2-fluoro-5-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (4 mg, 7.66 μmol). ¹H NMR in CD₃OD (400 MHz, METHANOL-d₄) δ7.03-6.98 (m, 3H), 6.95 (t, J=9.2 Hz, 1H), 6.80 (dd, J=6.2, 3.1 Hz, 1H),6.73 (dt, J=8.8, 3.5 Hz, 1H), 3.77 (s, 3H), 3.71-3.56 (m, 2H), 3.17-3.04(m, 1H), 2.90 (dd, J=16.6, 4.3 Hz, 1H), 2.85-2.68 (m, 4H), 2.52-2.35 (m,2H), 2.20-2.08 (m, 1H), 2.07-1.88 (m, 4H), 1.82-1.59 (m, 4H), 1.44 (dtd,J=12.8, 10.4, 6.1 Hz, 1H). MS (m+1)=398. HPLC Peak RT=8.01 min.(Condition L) Purity=98%.

The Examples in Table 17 were prepared according to the generalprocedure of Example 305.

TABLE 17 HPLC ret. Ex. Time HPLC MS No. Structure MW (min.) condition(M⁺¹) 306

397.5 7.99 L 398 307

397.5 7.70 L 398 308

397.5 8.07 L 398 309

393.6 8.36 L 394 310

393.6 8.38 L 394 311

393.6 8.33 L 394 312

393.6 1.83 A 393 313

397.5 9.04 L 398

Example 314((1R,3S)-1-amino-3-((S)-6-(5-methoxy-5-methylhexyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a mixture of(5R,7S)-7-((S)-6-(5-methylhex-4-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(20 mg, 0.054 mmol) in MeOH (10 mL) was added mercuric acetate (26.0 mg,0.082 mmol). After 1 hour, LCMS showed almost complete conversion to newpeak that had the mass of desired product as the Hg adduct. A solutionof sodium borohydride (10.29 mg, 0.272 mmol) in sodium hydroxide (0.5mL, 0.500 mmol) was added to the reaction mixture to remove Hg. Themixture was filtered to remove solids. Next, additional 1N NaOH wasadded to the filtrate and the mixture was heated to 95° C. overnight.The mixture was cooled and acidified with TFA then purified by HPLC.HPLC conditions: Phenomenex Luna 5 micron C18 column (30×100 mm); MeCN(0.1% TFA)/water (0.1% TFA); 20%-100% gradient over 15 minutes; 30mL/min. Isolated fractions with correct mass and freeze-dried overnightto afford((1R,3S)-1-amino-3-((S)-6-(5-methoxy-5-methylhexyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (10 mg, 0.019 mmol). HPLC Peak RT=7.62 min (Condition L) MS(m+1)=374. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.03-6.96 (m, 3H), 3.72-3.55(m, 2H), 3.20 (s, 3H), 3.15-3.04 (m, 1H), 2.89-2.73 (m, 3H), 2.48-2.31(m, 2H), 2.19-2.05 (m, 1H), 2.03-1.88 (m, 4H), 1.81-1.62 (m, 2H),1.59-1.49 (m, 2H), 1.48-1.29 (m, 7H), 1.17 (s, 6H).

Example 315((1R,3S)-1-amino-3-((R)-6-(4-isopropoxybutyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a stirred solution of4-((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)butanal(27 mg, 0.079 mmol), isoproxytrimethylsilane (0.070 mL, 0.395 mmol), andtriethylsilane (0.063 mL, 0.395 mmol) in nitromethane (1 mL) was addedferric chloride (1.283 mg, 7.91 μmol) at 0° C. under nitrogen. Themixture was stirred at 0° C. for 15 min and at room temperature for 30min. The mixture was concentrated. The residue was mixed with saturatedaqueous sodium bicarbonate solution (1 mL) and extracted with ethylacetate (3×1 mL). The combined ethyl acetate extracts were dried(Na₂SO₄) and concentrated under reduced pressure. The crude product wasdissolved in MeOH/DMSO (1:1) and treated with 1N NaOH at 95° C.overnight. LCMS show complete hydrolysis. The mixture was acidified withTFA then filtered and purified by HPLC. HPLC conditions: Phenomenex Luna5 micron C18 column (30×100 mm); MeCN (0.1% TFA)/water (0.1% TFA);20%-100% gradient over 15 minutes; 30 mL/min. Fractions with the correctmass were isolated and freeze-dried overnight to afford((1R,3S)-1-amino-3-((R)-6-(4-isopropoxybutyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (25 mg, 0.048 mmol). MS (m+1)=360. HPLC Peak RT=7.48 min (ConditionL). ¹H NMR (400 MHz, METHANOL-d₄) δ 7.03-6.95 (m, 3H), 3.74-3.56 (m,3H), 3.48 (t, J=6.4 Hz, 2H), 3.18-3.03 (m, 1H), 2.93-2.71 (m, 3H),2.49-2.31 (m, 2H), 2.20-2.05 (m, 1H), 2.03-1.87 (m, 4H), 1.73 (t, J=12.8Hz, 2H), 1.58 (q, J=6.5 Hz, 2H), 1.54-1.45 (m, 2H), 1.45-1.30 (m, 3H),1.17 (d, J=6.2 Hz, 6H).

Example 316((1R,3S)-1-amino-3-((6S)-6-(5-methoxyhexyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a mixture of(5R,7S)-7-((S)-6-(hex-5-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(38 mg, 0.107 mmol) in MeOH (1 mL) was added mercuric acetate (34.3 mg,0.107 mmol). The reaction mixture was stirred for 2 h then checked byLCMS. LCMS showed desired product mass plus Hg. A solution of sodiumborohydride (20.33 mg, 0.537 mmol) in 1M sodium hydroxide (1.075 mL,1.075 mmol) was added. The mixture was stirred for one hour. The mixturewas filtered to remove solids. The filtrate was then heated in 1NNaOH/MeOH at 95° C. overnight, cooled and acidified with TFA, and thenpurified by HPLC. HPLC conditions: Phenomenex Luna 5 micron C18 column(30×100 mm); MeCN (0.1% TFA)/water (0.1% TFA); 20%-100% gradient over 15minutes; 30 mL/min. Fractions with the correct mass were isolated andfreeze-dried overnight to afford((1R,3S)-1-amino-3-((6S)-6-(5-methoxyhexyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol, TFA (18 mg, 0.037 mmol). HPLC Peak RT=7.69/min (Condition L)MS (m+1)=360. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.03-6.96 (m, 3H),3.72-3.56 (m, 2H), 3.39-3.35 (m, 1H), 3.34 (s, 3H), 3.18-3.03 (m, 1H),2.91-2.74 (m, 3H), 2.49-2.30 (m, 2H), 2.18-2.05 (m, 1H), 2.03-1.87 (m,4H), 1.80-1.64 (m, 2H), 1.56 (d, J=3.7 Hz, 1H), 1.50-1.27 (m, 8H), 1.15(d, J=6.2 Hz, 3H).

Examples 317 to 322(1-amino-3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopentyl)methanol

Preparation 317A: 8-hexylidene-1,4-dioxaspiro[4.5]decane

To a mixture of hexyltriphenylphosphonium, iodide salt (25.6 g, 54 mmol)in THE (100 mL) was added LiHMDS (60 mL, 60.0 mmol). The reactionmixture was stirred for 15 minutes, then 1,4-dioxaspiro[4.5]decan-8-one(8.43 g, 54.0 mmol) in THF (100 mL) was added dropwise. The reactionmixture was stirred overnight. The reaction mixture was diluted withethyl acetate and washed with saturated NaCl. The organic layer wasdried with MgSO₄, filtered, and concentrated. The crude material waspurified on a silica gel cartridge (120 g) using an EtOAc/Hex gradient(100% hexanes for 4 CV then 0-30% EtOAc over 6 CV). Isolated fractionswith desired product, concentrated and dried in vacuo. Recovered 3.5 gof 8-hexylidene-1,4-dioxaspiro[4.5]decane.

Preparation 317B: 4-hexylcyclohexanone

To a mixture of 8-hexylidene-1,4-dioxaspiro[4.5]decane (3.5 g, 15.60mmol) in MeOH (30 mL) was added Pearlman's Catalyst (0.219 g, 1.560mmol). The reaction mixture was hydrogenated at 50 psi for 2 hours. Themixture was filtered and concentrated. Residue was dissolved in acetoneand treated with 1N HCl (20 ml of each). After stirring for 1 hour, thereaction mixture was diluted with ethyl acetate and washed withsaturated NaCl. The organic layer was dried with MgSO₄, filtered andconcentrated to afford 4-hexylcyclohexanone (2.8 g, 15.36 mmol).

Preparation 317C: 6-hexyl-5,6,7,8-tetrahydroquinolin-2-ol

To a mixture of 4-hexylcyclohexanone (2.8 g, 15.36 mmol), pyrrolidine(1.397 mL, 16.89 mmol), and p-toluenesulfonic acid monohydrate (0.088 g,0.461 mmol) in toluene (100 mL) was added molecular sieves. The reactionmixture was heated at 100° C. overnight. The mixture was filtered andthe solvent was removed. This material was dissolved in MeOH (30 mL) ina stainless steel pressure vessel. The vessel was cooled to −78° C. andammonia was bubbled in for 10 minutes. Methyl propriolate (3.87 mL, 46.1mmol) was added and the vessel was sealed and heated at 100° C. for 4hours. The reaction mixture was cooled in an ice bath and then ventedand opened. The reaction mixture was diluted with ethyl acetate andwashed with water. The organic layer was dried with MgSO₄, filtered andconcentrated. The crude material was purified on a silica gel cartridge(80 g) using an 20% MeOH/DCM:DCM gradient (0-50% 20% MeOH/DCM over 15CV). Product containing fractions were combined, concentrated and driedin vacuo to afford 6-hexyl-5,6,7,8-tetrahydroquinolin-2-ol (2.5 g, 10.71mmol).

Preparation 317D: 2-bromo-6-hexyl-5,6,7,8-tetrahydroquinoline (Isomers 1and 2)

To a mixture of 6-hexyl-5,6,7,8-tetrahydroquinolin-2-ol (580 mg, 2.486mmol) and phosphorus tribromide (4.97 mL, 4.97 mmol) in toluene (5 mL)was added phosphorus oxybromide (713 mg, 2.486 mmol). The reactionmixture was heated at 100° C. for 3 days. The mixture was cooled to 0°C. and then poured onto ice. The reaction mixture was diluted with ethylacetate and washed with saturated NaHCO₃. The organic layer was driedwith MgSO₄, filtered and concentrated. The crude material was purifiedon a silica gel cartridge (40 g) using an EtOAc/Hex gradient (0-50%EtOAc over 12 CV). Product containing fractions were combined,concentrated and dried in vacuo to afford2-bromo-6-hexyl-5,6,7,8-tetrahydroquinoline (300 mg, 1.013 mmol).

Preparations 317E1 and 317E2:3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopent-2-enone (Isomers 1and 2)

To a mixture of 2-bromo-6-hexyl-5,6,7,8-tetrahydroquinoline (885 mg,2.99 mmol) in THF (5 mL) was added n-BuLi (2.80 mL, 4.48 mmol) dropwise.The reaction mixture was stirred for 30 minutes. Next,3-ethoxycyclopent-2-enone (1.774 mL, 14.94 mmol) and lanthanum chloride(1465 mg, 5.97 mmol) were added. The reaction mixture was allowed towarm to 0° C. After 3 hours, the reaction was quenched with water. Thereaction mixture was diluted with ethyl acetate and washed withsaturated NaCl. The organic layer was dried with MgSO₄, filtered, andconcentrated. The crude material was purified on a silica gel cartridge(80 g) using an EtOAc/Hex gradient (0-30% EtOAc over 20 CV). The productcontaining fractions were combined, concentrated and dried in vacuo toafford 440 mg of3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopent-2-enone. ¹H NMR(400 MHz, CHLOROFORM-d) δ 7.57-7.42 (m, 2H), 6.85 (s, 1H), 3.18 (dd,J=5.0, 2.5 Hz, 3H), 3.07-2.86 (m, 2H), 2.62 (dt, J=5.0, 2.4 Hz, 2H),2.49 (dd, J=16.9, 10.3 Hz, 1H), 2.16-2.02 (m, 1H), 1.80 (br. s., 1H),1.54 (dtd, J=13.2, 11.0, 5.5 Hz, 1H), 1.42 (br. s., 4H), 1.33 (br. s.,6H), 1.00-0.82 (m, 3H). The isomers were separated by SFC using aChiralpak AD-H, 25×3 cm ID, 5 μm column and eluting with 70/30 CO₂/MeOHat 85.0 mL/min. Recovered two fractions which were concentrated anddried in vacuo. Isomer 1: Recovered3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopent-2-enone (210 mg,0.706 mmol). Isomer 2: Recovered3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopent-2-enone (210 mg,0.706 mmol).

Preparation 317F1 and 317F2

To a mixture of3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopent-2-enone (210 mg,0.706 mmol) (Isomer 1; Preparation 317E1) in MeOH (10 mL) and aceticacid (1 mL) was added Pearlman's Catalyst (50 mg, 0.356 mmol). Thereaction mixture was hydrogenated under a balloon of H₂. After 3 hours,the reaction mixture was filtered and concentrated in vacuo. The isomerswere separated by SFC using a Chiralpak IA-H, 25×2.1 cm ID, 5 μm columnand eluting with 95/5 CO₂/MeOH-ACN 1-1 at 50.0 mL/min. Recovered twofractions which were concentrated and dried in vacuo. Isomer 1A;recovered 45 mg; NMR was consistent with desired product ¹H NMR (400MHz, CHLOROFORM-d) δ 7.37-7.25 (m, 1H), 6.94 (d, J=7.7 Hz, 1H),3.61-3.43 (m, 1H), 3.04-2.76 (m, 3H), 2.74-2.53 (m, 2H), 2.52-2.21 (m,4H), 2.21-2.08 (m, 1H), 2.08-1.95 (m, 1H), 1.87-1.60 (m, 2H), 1.58-1.44(m, 1H), 1.44-1.21 (m, 9H), 1.01-0.81 (m, 3H). Isomer 1B; recovered 33mg; NMR was consistent with desired product. ¹H NMR (400 MHz,CHLOROFORM-d) δ 7.31 (d, J=7.7 Hz, 1H), 6.94 (d, J=7.9 Hz, 1H),3.60-3.44 (m, 1H), 3.03-2.75 (m, 3H), 2.71-2.54 (m, 2H), 2.54-2.34 (m,3H), 2.34-2.22 (m, 1H), 2.22-2.09 (m, 1H), 2.09-1.98 (m, 1H), 1.85-1.66(m, 2H), 1.60-1.44 (m, 1H), 1.44-1.23 (m, 9H), 0.97-0.86 (m, 3H).

Preparation 317G1 and 317G2: methyl1-amino-3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopentanecarboxylate(Isomers 1 and 2)

To a mixture of3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopentanone (45 mg, 0.150mmol), ammonium chloride (40.2 mg, 0.751 mmol), and sodium cyanide (36.8mg, 0.751 mmol) in DCM (5 mL) was added ammonia in MeOH (0.429 mL, 3.01mmol). The reaction mixture was sealed and stirred for 3 days. Thereaction was incomplete as indicated by LCMS analysis. Additional sodiumcyanide (36.8 mg, 0.751 mmol) and ammonium chloride (40.2 mg, 0.751mmol) were added and the reaction mixture was stirred for an additionalday. LCMS showed reaction was complete. The reaction mixture was dilutedwith dichloromethane and washed with water. The organic layer was driedwith MgSO₄, filtered and concentrated. The crude product was dissolvedin dioxane (1 mL), then acetic acid (1 mL) and concentrated HCl (1 mL)were added. The reaction mixture was heated at 100° C. overnight. Thereaction mixture was concentrated to dryness then crude material wasdissolved in MeOH. HCl (g) was bubbled through for 5 minutes. Themixture was heated at 70° C. for 1 hour. LCMS showed conversion to thedesired methyl ester. The mixture was concentrated in vacuo and purifiedby HPLC. HPLC conditions: Phenomenex Luna 5 micron C18 column (30×100mm); MeCN (0.1% TFA)/water (0.1% TFA); 20%-100% gradient over 15minutes; 30 mL/min. Isolated fractions with correct mass andfreeze-dried overnight. Recovered methyl1-amino-3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopentanecarboxylate,TFA (37 mg, 0.078 mmol). ¹H NMR (400 MHz, METHANOL-d₄) δ 8.33-8.06 (m,1H), 7.91-7.62 (m, 1H), 3.93 (s, 3H), 3.90-3.79 (m, 1H), 3.28-2.98 (m,3H), 2.90 (dd, J=13.9, 7.7 Hz, 1H), 2.79-2.40 (m, 4H), 2.38-2.08 (m,3H), 1.85 (br. s., 1H), 1.69-1.51 (m, 1H), 1.52-1.24 (m, 10H), 1.06-0.83(m, 3H). The isomers were separated by SFC using a Chiralpak OZ-H, 25×3cm ID, 5 μm column and eluting with 65/35 CO₂/MeOH w/0.1% DEA at 85.0mL/min. Two fractions were recovered which were concentrated and driedin vacuo. Isomer 1: methyl1-amino-3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopentanecarboxylate,TFA (15 mg, 0.032 mmol). Isomer 2: methyl1-amino-3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopentanecarboxylate(18 mg, 0.050 mmol).

Example 317

To a mixture of methyl1-amino-3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopentanecarboxylate, TFA (15 mg, 0.032 mmol) (Isomer 1; Preparation317G1) in MeOH (3 mL) was added sodium borohydride (7.21 mg, 0.190mmol). After 2 hour, the reaction was quenched with water. The reactionmixture was concentrated and the residue was triturated in TFA/MeCN, andthen filtered. The filtrate was purified by HPLC. HPLC conditions:Phenomenex Luna 5 micron C18 column (30×100 mm); MeCN (0.1% TFA)/water(0.1% TFA); 10%-100% gradient over 15 minutes; 30 mL/min. Isolatedfractions with correct mass and freeze-dried overnight to afford(1-amino-3-(6-hexyl-5,6,7,8-tetrahydroquinolin-2-yl)cyclopentyl)methanol,2 TFA (12.6 mg, 0.021 mmol). ¹H NMR in CD₃OD was consistent with desiredproduct (400 MHz, METHANOL-d₄) δ 8.21 (d, J=8.1 Hz, 1H), 7.73 (d, J=8.1Hz, 1H), 3.80 (ddd, J=10.7, 7.5, 3.3 Hz, 1H), 3.75-3.63 (m, 2H),3.28-3.01 (m, 3H), 2.56 (dd, J=17.2, 10.6 Hz, 1H), 2.48-2.25 (m, 3H),2.22-2.08 (m, 2H), 2.07-1.89 (m, 2H), 1.89-1.77 (m, 1H), 1.59 (dtd,J=13.3, 11.0, 5.8 Hz, 1H), 1.47 (d, J=3.1 Hz, 4H), 1.36 (d, J=3.1 Hz,6H), 1.00-0.88 (m, 3H); HPLC retention time=6.81 min (condition L);LC/MS M⁺¹=331.

The Examples 318-322 in Table 18 were prepared according to the generalprocedure of Example 317.

TABLE 18 HPLC ret. Ex. Time HPLC MS No. Structure MW (min.) condition(M⁺¹) Comments 318 319 320 321     322

330.5 330.5 330.5 330.5     330.5 6.73 6.80 6.73 6.95     6.95 L L L L    L 331 331 331 331     331 Isomer 1A2 Isomer 1B1 Isomer 1B2 Isomer 2AMixture of 2 diastereomers Isomer 2B Mixture of 2 diastereomers

Examples 326 to 3295-(3-amino-3-(hydroxymethyl)cyclopentyl)-2-(3-phenylpropyl)isoindolin-1-one

Preparation 326A: 6-bromo-2-(pentyloxy)quinoline

To a solution of methyl 4-bromo-2-(bromomethyl)benzoate (2.000 g, 6.49mmol) and 3-phenyl-1-propylamine (1.016 mL, 7.14 mmol) in EtOH (15 mL)was added potassium carbonate (1.346 g, 9.74 mmol). The reaction mixturewas heated at 40° C. for 3 h. The reaction mixture was diluted withethyl acetate and washed with saturated NaCl. The organic layer wasdried with MgSO₄, filtered and concentrated. The crude oil was purifiedon a 80 g silica gel cartridge using 30-60% EtOAc/hexanes gradient toafford 5-bromo-2-(3-phenylpropyl)isoindolin-1-one (1.43 g, 4.33 mmol) asa white solid. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.77-7.71 (m, 1H),7.66-7.59 (m, 2H), 7.35-7.26 (m, 2H), 7.24-7.15 (m, 3H), 4.35 (s, 2H),3.69 (t, J=7.3 Hz, 2H), 2.79-2.66 (m, 2H), 2.13-1.96 (m, 2H).

Examples 326 to 329

An oven dried microwave vial with a stir bar was charged with5-bromo-2-(3-phenylpropyl)isoindolin-1-one (750 mg, 2.271 mmol), ethyl1-((diphenylmethylene) amino)cyclopent-3-enecarboxylate (1233 mg, 3.86mmol), palladium(II) acetate (102 mg, 0.454 mmol), triphenylphosphine(238 mg, 0.908 mmol), potassium acetate (446 mg, 4.54 mmol) and DMA (20mL). The mixture was sparged with nitrogen for 10 minutes. The solutionwas processed on a CEM microwave: 60 minutes at 140° C. The reactionmixture was diluted with ethyl acetate and washed with saturated NaCl.The organic layer was dried with MgSO₄, filtered and concentrated. Thecrude material was purified on a silica gel cartridge (80 g) using anEtOAc/Hex gradient (0-100% EtOAc over 20 minutes) to afford 825 mg ofmaterial. This residue was dissolved in ether (20 mL) and treated with6N HCl for 30 minutes. The reaction mixture was diluted with ethylacetate and washed with saturated NaHCO₃. The organic layer was driedwith MgSO₄, filtered and concentrated. This residue was dissolved inethanol (20 mL) and sodium borohydride (859 mg, 22.71 mmol) was addedportionwise over several hours until no starting material remained. Thereaction was quenched with 1N HCl. The reaction mixture was diluted withethyl acetate and washed with saturated NaHCO₃. The organic layer wasdried with MgSO₄, filtered and concentrated. This residue was dissolvedin MeOH and 10% Pd/C was added. The reaction mixture was hydrogenatedunder a balloon of H₂ for 1 hour. The reaction mixture was filtered andpurified by HPLC. HPLC conditions: Phenomenex Luna 5 micron C18 column(30×100 mm); MeCN (0.1% TFA)/water (0.1% TFA); 30%-100% gradient over 10minutes; 30 mL/min. Fractions with correct mass were isolated, dilutedwith ethyl acetate, washed with saturated NaHCO₃, and back extractedtwice with EtOAc. The organic layer was dried with MgSO₄, filtered andconcentrated to afford 275 mg of 5-(3-amino-3-(hydroxymethyl)cyclopentyl)-2-(3-phenylpropyl)isoindolin-1-one. The individual isomerswere separated using a CHIRALPAK® AD-H column under SFC conditions (20%MeOH with 0.5% DEA in CO₂).

Example 326 (33 mg) HPLC retention time=5.55 min (condition H); LC/MSM⁺¹=X; ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.78 (d, J=7.9 Hz, 1H),7.38-7.27 (m, 5H), 7.25-7.16 (m, 3H), 4.34 (s, 2H), 3.68 (t, J=7.3 Hz,2H), 3.52 (d, J=5.9 Hz, 2H), 2.16-1.87 (m, 8H), 1.83-1.66 (m, 2H),1.66-1.52 (m, 1H).

Example 327 (105 mg) HPLC retention time=5.61 min (condition H); LC/MSM⁺¹=X; ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.77 (d, J=8.4 Hz, 1H),7.40-7.34 (m, 2H), 7.33-7.26 (m, 2H), 7.25-7.16 (m, 3H), 4.34 (s, 2H),3.68 (t, J=7.3 Hz, 2H), 3.56 (br. s., 2H), 3.20 (t, J=7.5 Hz, 1H),2.77-2.61 (m, 2H), 2.45-2.31 (m, 1H), 2.21-2.08 (m, 1H), 2.08-1.90 (m,3H), 1.83 (br. s., 2H), 1.65 (t, J=12.0 Hz, 1H).

Example 328 (25 mg) HPLC retention time=5.47 min (condition H); LC/MSM+¹ ═X; ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.78 (d, J=7.9 Hz, 1H),7.38-7.26 (m, 4H), 7.25-7.17 (m, 3H), 4.34 (s, 2H), 3.68 (t, J=7.3 Hz,2H), 3.52 (d, J=5.9 Hz, 2H), 3.36 (dd, J=10.1, 4.2 Hz, 1H), 2.80-2.63(m, 2H), 2.14-1.91 (m, 5H), 1.88-1.67 (m, 2H), 1.62 (dd, J=12.7, 5.2 Hz,1H).

Example 329 (100 mg) HPLC retention time=5.60 min (condition H); LC/MSM⁺¹=X; ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.77 (d, J=7.9 Hz, 1H),7.39-7.33 (m, 2H), 7.31-7.25 (m, 2H), 7.23-7.14 (m, 3H), 4.33 (s, 2H),3.67 (t, J=7.3 Hz, 2H), 3.48 (d, J=8.6 Hz, 2H), 3.19 (t, J=7.6 Hz, 1H),2.34 (dd, J=12.8, 8.1 Hz, 1H), 2.21-1.88 (m, 6H), 1.87-1.64 (m, 2H),1.54 (t, J=11.8 Hz, 1H).

The absolute stereochemistries of the isomers were not determined.

Examples 330 to 3325-(3-amino-3-(hydroxymethyl)cyclopentyl)-3,3-dimethyl-2-(3-phenylpropyl)isoindolin-1-one

Preparation 330A: 5-methoxy-3,3-dimethylisoindolin-1-one

To a mixture of 2-(3-methoxyphenyl)-2-methylpropanoic acid (13.01 g, 67mmol) and Et₃N (9.34 mL, 67.0 mmol) in toluene (200 mL) at 0° C. wasadded diphenylphosphoryl azide (14.40 mL, 67.0 mmol). After 30 min at 0°C., the reaction mixture was warmed to room temperature then refluxedovernight. The reaction mixture was diluted with ethyl acetate andwashed with saturated NaHCO₃ and brine. The organic layer was dried withMgSO₄, filtered and concentrated. This crude residue was dissolved inDCE (100 mL) and added dropwise to a slurry of iron(III) chloride (23.91g, 147 mmol) in DCE (300 mL) at 0° C. The mixture was stirred for 2hours and allowed to warm to room temperature. The reaction mixture wasdiluted with 1M tartaric acid solution and stirred for 30 minutes. Theorganic layer was separated then dried with MgSO₄, filtered andconcentrated. The crude material was purified on a silica gel cartridge(40 g) using an EtOAc/Hex gradient (40-100% EtOAc over 11 CV then heldat 100% EtOAc until product completely eluted) to afforded 4.9 g of5-methoxy-3,3-dimethylisoindolin-1-one. HPLC retention time=0.87 min(condition G); LC/MS M⁺¹=192; ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.74(1H, d, J=7.70 Hz), 7.40 (1H, br. s.), 6.97 (1H, d, J=7.48 Hz), 6.87(1H, br. s.), 3.90 (3H, br. s.), 1.55 (6H, br. s.).

Preparation 330B:5-methoxy-3,3-dimethyl-2-(3-phenylpropyl)isoindolin-1-one

To a mixture of 5-methoxy-3,3-dimethylisoindolin-1-one (1.8 g, 9.41mmol) in DMF (50 mL) was added sodium hydride (0.565 g, 14.12 mmol)portionwise. After addition, the reaction mixture was heated to 80° C.for 1 hour followed by the addition of 3-iodopropyl)benzene (3.03 mL,18.83 mmol). Reaction was incomplete after 2 hours.

Additional sodium hydride (0.565 g, 14.12 mmol) was added and thereaction mixture was heated overnight. Reaction was still incomplete.Additional sodium hydride (0.565 g, 14.12 mmol) was added and heatingwas continued for 4 more hours. The reaction mixture was diluted withethyl acetate and washed twice with saturated NaCl. The organic layerwas dried with MgSO₄, filtered and concentrated. The crude material waspurified on a silica gel cartridge (40 g) using an EtOAc/Hex gradient(0-100% EtOAc over 13 CV) to afford 850 mg of5-methoxy-3,3-dimethyl-2-(3-phenylpropyl)isoindolin-1-one. HPLCretention time=1.00 min (condition G); LC/MS M⁺¹=310; ¹H NMR (400 MHz,CHLOROFORM-d) δ 7.75 (d, J=8.4 Hz, 1H), 7.46-7.14 (m, 5H), 6.96 (dd,J=8.4, 2.2 Hz, 1H), 6.86 (d, J=2.0 Hz, 1H), 3.90 (s, 3H), 2.75 (td,J=7.8, 5.4 Hz, 2H), 2.16-2.02 (m, 2H), 2.00-1.86 (m, 2H), 1.60 (s, 3H),1.46 (s, 3H). MS (m+1)=310.

Preparation 330C: 55-hydroxy-3,3-dimethyl-2-(3-phenylpropyl)isoindolin-1-one

To a mixture of5-methoxy-3,3-dimethyl-2-(3-phenylpropyl)isoindolin-1-one (850 mg, 2.75mmol) in DCM (Volume: 10 mL) was added BBr₃ in DCM (5.49 mL, 5.49 mmol).The reaction mixture was heated at 50° C. for 5 hours. The reactionmixture was diluted with DCM and washed with saturated NaHCO₃. Theorganic layer was dried with MgSO₄, filtered and concentrated. DCM wasadded and solid material precipitated. The mixture was allowed to in arefrigerator for 1 hour. The solid was collected by filtration and driedto afford 450 mg of5-hydroxy-3,3-dimethyl-2-(3-phenylpropyl)isoindolin-1-one as a tansolid. HPLC retention time=0.87 min (condition G); LC/MS M⁺¹=296; ¹H NMR(400 MHz, DMSO-d₆) δ ppm 10.08 (1H, s), 7.43 (1H, d, J=8.14 Hz),7.13-7.35 (5H, m), 6.92 (1H, d, J=1.98 Hz), 6.82 (1H, dd, J=8.25, 2.09Hz), 3.36-3.41 (2H, m), 2.60-2.73 (2H, m), 1.80-1.98 (2H, m), 1.39 (6H,s). MS (m+1)=295.

Preparation 330D: 3,3-dimethyl-1-oxo-2-(3-phenylpropyl)isoindolin-5-yltrifluoromethanesulfonate

To a mixture of5-hydroxy-3,3-dimethyl-2-(3-phenylpropyl)isoindolin-1-one (440 mg, 1.490mmol) and pyridine (361 μl, 4.47 mmol) in DCM was added triflicanhydride (377 μl, 2.234 mmol). The reaction mixture was stirred for onehour. The reaction mixture was diluted with DCM and washed withsaturated NaCl. The organic layer was dried with MgSO₄, filtered andconcentrated to afford 600 mg of3,3-dimethyl-1-oxo-2-(3-phenylpropyl)isoindolin-5-yltrifluoromethanesulfonate which was used immediately in the next step.HPLC retention time=1.09 min (condition G); LC/MS M⁺¹=428.

Preparation 330E: Ethyl4-(3,3-dimethyl-1-oxo-2-(3-phenylpropyl)isoindolin-5-yl)-1-(diphenylmethyleneamino)cyclopent-2-enecarboxylate

An oven dried microwave vial with stir bar was charged with3,3-dimethyl-1-oxo-2-(3-phenylpropyl)isoindolin-5-yltrifluoromethanesulfonate (662 mg, 1.550 mmol), ethyl1-((diphenylmethylene)amino)cyclopent-3-enecarboxylate (330 mg, 1.033mmol), palladium(II) acetate (46.4 mg, 0.207 mmol), triphenylphosphine(108 mg, 0.413 mmol), potassium acetate (203 mg, 2.066 mmol) and DMA (4mL). The mixture was sparged with nitrogen for 10 minutes. The solutionwas processed on a CEM microwave: 60 minutes at 140° C. The reactionmixture was diluted with ethyl acetate and washed with saturated NaCl.The organic layer was dried with MgSO₄, filtered and concentrated. Thecrude material was purified on a silica gel cartridge (80 g) using anEtOAc/Hex gradient (0-100% EtOAc over 20 minutes) to afford 330 mg ofethyl4-(3,3-dimethyl-1-oxo-2-(3-phenylpropyl)isoindolin-5-yl)-1-(diphenylmethyleneamino)cyclopent-2-enecarboxylate.HPLC retention time=1.05 min (condition G); LC/MS M⁺¹=597.

Examples 330 to 332

To a mixture of ethyl4-(3,3-dimethyl-1-oxo-2-(3-phenylpropyl)isoindolin-5-yl)-1-((diphenylmethylene)amino)cyclopent-2-enecarboxylate(330 mg, 0.553 mmol) in ether (10 mL) was added 6N HCl (5 mL). Thereaction mixture was stirred for 30 minutes. The reaction mixture wasdiluted with ethyl acetate and washed with saturated NaHCO₃. The organiclayer was dried with MgSO₄, filtered and concentrated. This residue wasdissolved in MeOH (10.00 mL) and sodium borohydride (105 mg, 2.76 mmol)was added. Additional sodium borohydride (105 mg, 2.76 mmol) was addeduntil LCMS showed complete conversion of the starting material. Thereaction was quenched with 1N HCl then the reaction mixture was dilutedwith ethyl acetate and washed with saturated NaHCO₃. The organic layerwas dried with MgSO₄, filtered and concentrated. This residue wasdissolved in MeOH and Pd/C (58.8 mg, 0.553 mmol) was added. The reactionmixture was hydrogenated under a balloon of H₂ for 1 hour, and thenfiltered and purified by HPLC. HPLC conditions: Phenomenex Luna 5 micronC18 column (30×100 mm); MeCN (0.1% TFA)/water (0.1% TFA); 20%-100%gradient over 15 minutes; 30 mL/min. Recovered 100 mg of5-(3-amino-3-(hydroxymethyl)cyclopentyl)-3,3-dimethyl-2-(3-phenylpropyl)isoindolin-1-one. Theindividual isomers were separated using a CHIRALPAK® AS-H column underSFC conditions (15% MeOH/IPA (1:1) with 0.5% DEA in CO₂).

Example 330: Fraction 1 (4 mg, mixture of two isomers) HPLC retentiontime=6.98 min (condition H); LC/MS M⁺¹=393; ¹H NMR (400 MHz,METHANOL-d₄) δ 7.77-7.64 (m, 1H), 7.49 (s, 1H), 7.47-7.38 (m, 1H),7.35-7.23 (m, 4H), 7.22-7.14 (m, 1H), 3.81-3.61 (m, 2H), 3.55-3.45 (m,3H), 2.74 (t, J=7.8 Hz, 2H), 2.36-2.18 (m, 2H), 2.11-2.00 (m, 3H),1.99-1.76 (m, 3H), 1.50 (s, 6H).

Example 331: Fraction 2 (13 mg, homochiral) HPLC retention time=7.02 min(condition H); LC/MS M⁺¹=393; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.67 (d,J=7.9 Hz, 1H), 7.51 (s, 1H), 7.48-7.39 (m, 1H), 7.34-7.24 (m, 4H),7.23-7.11 (m, 1H), 3.60-3.42 (m, 4H), 3.24 (ddd, J=11.2, 7.1, 4.0 Hz,1H), 2.74 (t, J=7.7 Hz, 2H), 2.31 (dd, J=13.0, 7.7 Hz, 1H), 2.18-1.97(m, 4H), 1.92-1.71 (m, 2H), 1.68-1.56 (m, 1H), 1.49 (s, 6H), MS(m+1)=393.

Example 332: Fraction 3 (17 mg, homochiral) HPLC retention time=6.99 min(condition H); LC/MS M⁺¹=393; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.67 (d,J=7.7 Hz, 1H), 7.51 (s, 1H), 7.43 (dd, J=7.9, 1.1 Hz, 1H), 7.33-7.23 (m,4H), 7.22-7.15 (m, 1H), 3.57-3.44 (m, 4H), 3.23 (ddd, J=10.9, 7.4, 3.7Hz, 1H), 2.74 (t, J=7.8 Hz, 2H), 2.31 (dd, J=13.1, 7.8 Hz, 1H),2.15-1.97 (m, 4H), 1.89-1.69 (m, 2H), 1.67-1.56 (m, 1H), 1.49 (s, 6H),MS (m+1)=393. The absolute stereochemistry of the isomers was notdetermined.

Examples 333 to 3351-(6-(3-amino-3-(hydroxymethyl)cyclopentyl)-3,4-dihydroisoquinolin-2(1H)-yl)hexan-1-one

Preparation 333A: 3-(isoquinolin-6-yl)cyclopentanone

To a mixture of 6-bromoisoquinoline (2 g, 9.61 mmol), cyclopent-2-enol(2.021 g, 24.03 mmol), and potassium acetate (2.83 g, 28.8 mmol) in DMF(50 mL) was added tetrabutylammonium chloride (2.67 g, 9.61 mmol) andpalladium (II) acetate (0.216 g, 0.961 mmol). The reaction mixture wasdegassed with nitrogen and then heated at 80° C. overnight. The reactionmixture was diluted with ethyl acetate and washed with saturated NaCl.The organic layer was dried with MgSO₄, filtered and concentrated. Thecrude material was purified on a silica gel cartridge (80 g) using anEtOAc/Hex gradient (20-100% EtOAc over 10 CV) to afford 750 mg of3-(isoquinolin-6-yl)cyclopentanone. HPLC retention time=0.52 min(condition H); LC/MS M⁺¹=212. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.92 (dd,J=4.2, 1.8 Hz, 1H), 8.22-8.08 (m, 2H), 7.74-7.61 (m, 2H), 7.43 (dd,J=8.4, 4.2 Hz, 1H), 3.65 (tt, J=10.7, 6.9 Hz, 1H), 2.80 (dd, J=18.3, 7.7Hz, 1H), 2.66-2.30 (m, 4H), 2.20-2.02 (m, 1H).

Preparation 333B:7-(isoquinolin-6-yl)-1,3-diazaspiro[4.4]nonane-2,4-dione

To a mixture of 3-(isoquinolin-6-yl)cyclopentanone (820 mg, 3.88 mmol)and potassium cyanide (379 mg, 5.82 mmol) in EtOH (20 mL) and water (10mL) in a pressure vessel was added potassium cyanide (379 mg, 5.82mmol). The vessel was sealed and heated at 90° C. overnight. Thereaction mixture was cooled and vented. The reaction mixture was dilutedwith ethyl acetate and washed with saturated NaCl. The organic layer wasdried with MgSO₄, filtered and concentrated to afford 890 mg of7-(isoquinolin-6-yl)-1,3-diazaspiro[4.4]nonane-2,4-dione. HPLC retentiontime=0.65 min (condition G); LC/MS M⁺¹=393. ¹H NMR (400 MHz, DMSO-d₆) δ10.65 (d, J=12.3 Hz, 1H), 8.86 (dd, J=4.2, 1.5 Hz, 1H), 8.40 (s, 1H),8.36-8.25 (m, 1H), 7.99 (d, J=8.6 Hz, 1H), 7.84 (d, J=2.2 Hz, 1H), 7.74(ddd, J=16.5, 8.7, 2.1 Hz, 1H), 7.52 (dd, J=8.4, 4.2 Hz, 1H), 3.65-3.39(m, 1H), 2.56 (dd, J=13.6, 8.1 Hz, 1H), 2.41-2.08 (m, 3H), 2.03-1.78 (m,2H).

Preparation 333C: methyl1-amino-3-(isoquinolin-6-yl)cyclopentanecarboxylate

To a mixture of 7-(isoquinolin-6-yl)-1,3-diazaspiro[4.4]nonane-2,4-dione(890 mg, 3.16 mmol) in MeOH (20 mL) was added 2N NaOH. After heating fortwo days, the reaction mixture was concentrated in vacuo and dried. Thecrude product was suspended in MeOH. HCl (g) was bubbled through for 15minutes then the reaction mixture was heated at 80° C. The solvent waspartially removed in vacuo, then the mixture was filtered and purifiedby HPLC. HPLC conditions: Phenomenex Luna C18 5 micron column (250×30mm); 10-100% MeCN/water (0.1% TFA); 25 minute gradient; 30 mL/min.Recovered 750 mg of methyl1-amino-3-(isoquinolin-6-yl)cyclopentanecarboxylate. HPLC retentiontime=0.43 min (condition G); LC/MS M⁺¹=271.

Preparation 333D: methyl1-(tert-butoxycarbonylamino)-3-(isoquinolin-6-yl)cyclopentanecarboxylate

To a mixture of methyl1-amino-3-(isoquinolin-6-yl)cyclopentanecarboxylate and DIEA (1.022 mL,5.85 mmol) in acetonitrile (10 mL) was added (Boc)₂₀ (1.359 mL, 5.85mmol). The reaction mixture was stirred at room temperature for 2 hours.The reaction mixture was diluted with ethyl acetate and washed withsaturated NaCl. The organic layer was dried with MgSO₄, filtered andconcentrated. The crude material was purified on a silica gel cartridge(24 g) using an EtOAc/Hex gradient (0-100% EtOAc over 12 CV). Recovered380 mg of methyl1-(tert-butoxycarbonylamino)-3-(isoquinolin-6-yl)cyclopentanecarboxylate.HPLC retention time=0.72 min (condition G); LC/MS M⁺¹=371. ¹H NMR (400MHz, CHLOROFORM-d) δ 8.88 (dd, J=4.2, 1.8 Hz, 1H), 8.09 (dd, J=18.3, 8.8Hz, 2H), 7.73-7.63 (m, 2H), 7.39 (dd, J=8.3, 4.3 Hz, 1H), 5.36-5.02 (m,1H), 3.81 (d, J=3.1 Hz, 3H), 3.68-3.43 (m, 1H), 2.67-2.25 (m, 3H),2.21-1.80 (m, 3H), 1.47 (d, J=5.1 Hz, 9H).

Preparation 333E: methyl1-(tert-butoxycarbonylamino)-3-(1,2,3,4-tetrahydroisoquinolin-6-yl)cyclopentanecarboxylate

To a mixture of methyl1-((tert-butoxycarbonyl)amino)-3-(isoquinolin-6-yl)cyclopentanecarboxylate (280 mg, 0.756 mmol) in acetic acid (10 mL) wasadded platinum(IV) oxide (17.16 mg, 0.076 mmol). The reaction mixturewas hydrogenated on a Parr shaker for 2 hours at 40 PSI of hydrogen. Thecatalyst was removed by filtration and the mixture was concentrated togive 200 mg of methyl1-(tert-butoxycarbonylamino)-3-(1,2,3,4-tetrahydroisoquinolin-6-yl)cyclopentanecarboxylate.HPLC retention time=0.70 min (condition G); LC/MS M⁺¹=375.

Preparation 333F: Methyl1-(tert-butoxycarbonylamino)-3-(2-hexanoyl-1,2,3,4-tetrahydroisoquinolin-6-yl)cyclopentanecarboxylate

To a mixture of methyl1-((tert-butoxycarbonyl)amino)-3-(1,2,3,4-tetrahydroisoquinolin-6-yl)cyclopentanecarboxylate(200 mg, 0.534 mmol) and DIEA (200 μl, 1.145 mmol) in DCM (5 mL) wasadded hexanoyl chloride (74.7 μl, 0.534 mmol). The reaction mixture wasstirred for 30 minutes. The reaction mixture was diluted with ethylacetate and washed with saturated NaCl. The organic layer was dried withMgSO₄, filtered and concentrated. The crude material was purified on asilica gel cartridge (24 g) using an EtOAc/Hex gradient (0-100% EtOAcover 13CV). Recovered 140 mg of methyl1-(tert-butoxycarbonylamino)-3-(2-hexanoyl-1,2,3,4-tetrahydroisoquinolin-6-yl)cyclopentanecarboxylate.HPLC retention time=1.12 min (condition G); LC/MS M⁺¹=473.

Examples 333 to 335

To a mixture of methyl1-((tert-butoxycarbonyl)amino)-3-(2-hexanoyl-1,2,3,4-tetrahydroisoquinolin-6-yl)cyclopentanecarboxylate(140 mg, 0.296 mmol) in DCM (2 mL) was added TFA (2 mL). The reactionmixture was stirred for 1 hour. LCMS shows complete removal of Bocgroup. The mixture was concentrated in vacuo, and MeOH (5 mL) was addedfollowed by portionwise addition of sodium borohydride (56.0 mg, 1.481mmol). After one hour, more sodium borohydride (112.0 mg, 3.5 mmol) wasadded. The reaction was quenched with water. The reaction mixture wasdiluted with ethyl acetate and washed with saturated NaCl. The organiclayer was dried with MgSO₄, filtered and concentrated. HPLC conditions:Phenomenex Luna 5 micron C18 column (30×100 mm); MeCN (0.1% TFA)/water(0.1% TFA); 10%-100% gradient over 15 minutes; 30 mL/min. Recovered 44mg of1-(6-(3-amino-3-(hydroxymethyl)cyclopentyl)-3,4-dihydroisoquinolin-2(1H)-yl)hexan-1-one.The individual isomers were separated using a CHIRALPAK® AS-H columnunder SFC conditions (15% MeOH with 0.1% DEA in CO₂).

Example 333: Isomer 1 (9 mg, racemic) HPLC retention time=6.60 min(condition H); LC/MS M⁺¹=393. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.17 (br.s., 3H), 3.77 (t, J=6.5 Hz, 2H), 3.58-3.43 (m, 2H), 3.10 (tt, J=11.3,7.2 Hz, 1H), 2.73 (t, J=6.5 Hz, 2H), 2.54 (t, J=7.5 Hz, 2H), 2.29 (dd,J=13.1, 7.6 Hz, 1H), 2.03-1.90 (m, 3H), 1.89-1.70 (m, 2H), 1.69-1.53 (m,3H), 1.42-1.16 (m, 5H), 0.97-0.81 (m, 3H).

Example 334: Isomer 2 (10 mg, homochiral) HPLC retention time=6.54 min(condition H); LC/MS M⁺¹=393. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.13 (br.s., 3H), 3.76 (t, J=6.6 Hz, 2H), 3.62-3.45 (m, 2H), 3.43-3.36 (m, 1H),2.85 (q, J=7.3 Hz, 1H), 2.72 (t, J=6.4 Hz, 2H), 2.54 (t, J=7.6 Hz, 2H),2.28-2.01 (m, 2H), 1.98-1.87 (m, 2H), 1.81-1.51 (m, 5H), 1.45-1.10 (m,6H), 0.89 (br. s., 3H).

Example 335: Isomer 3 (8.5 mg, homochiral) HPLC retention time=6.54 min(condition H); LC/MS M⁺¹=393. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.67 (d,J=7.7 Hz, 1H), 7.51 (s, 1H), 7.43 (dd, J=7.9, 1.1 Hz, 1H), 7.33-7.23 (m,4H), 7.22-7.15 (m, 1H), 3.57-3.44 (m, 4H), 3.23 (ddd, J=10.9, 7.4, 3.7Hz, 1H), 2.74 (t, J=7.8 Hz, 2H), 2.31 (dd, J=13.1, 7.8 Hz, 1H),2.15-1.97 (m, 4H), 1.89-1.69 (m, 2H), 1.67-1.56 (m, 1H), 1.49 (s, 6H),MS (m+1)=393. The absolute stereochemistry of the isomers was notdetermined.

Example 336(((1R,3S)-1-amino-3-((6S)-6-((phenylsulfinyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a stirred clear solution of((1R,3S)-1-amino-3-((S)-6-((phenylthio)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(6 mg, 0.016 mmol), DMSO (0.035 mL, 0.490 mmol), and L-10-(−)-camphorsulfonic acid (18.96 mg, 0.082 mmol) in dichloromethane (0.5 mL) andmethanol (0.2 mL) cooled with dry-ice was added 77% m-CPBA (3.66 mg,0.016 mmol). The temperature was raised to 0° C. over 30 min. Themixture was stirred at 0° C. for 30 min and room temperature for 30 min.The mixture was concentrated and purified using reverse phase HPLC(Waters Xbridge C18 19×100 mm; gradient over 8 min from 20 to 100% ofsolvent B; solvent A: 10% MeOH: 90% H₂O: 0.1% TFA; solvent B: 90% MeOH,10% H₂O, 0.1% TFA), concentration, basification with K₂CO₃, andextraction with ethyl acetate gave((1R,3S)-1-amino-3-((6S)-6-((phenylsulfinyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol (6 mg, 0.015 mmol) as a glassy solid. LC/MS M⁺¹=384. ¹H NMR(500 MHz, CHLOROFORM-d) δ 7.68-7.62 (m, 2H), 7.56-7.47 (m, 3H),7.03-6.94 (m, 3H), 3.50-3.41 (m, 2H), 3.18-2.90 (m, 3H), 2.87-2.77 (m,2H), 2.73-2.52 (m, 2H), 2.47-2.36 (m, 1H), 2.26 (dd, J=13.3, 7.8 Hz,1H), 2.21-1.97 (m, 2H), 1.96-1.82 (m, 1H), 1.79-1.70 (m, 1H), 1.70-1.58(m, 2H), 1.49 (dd, J=13.3, 11.1 Hz, 1H).

Example 337((1R,3S)-1-amino-3-((S)-6-((phenylsulfonyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a stirred solution of((1R,3S)-1-amino-3-((S)-6-((phenylthio)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(2 mg, 5.44 μmol) and L-10-(−)-camphor sulfonic acid (6.32 mg, 0.027mmol) in dichloromethane (5 mL) was added 77% m-CPBA (3.13 mg, 10.88μmol). The mixture was stirred at room temperature for 3 h.

Purification using reverse phase HPLC (Waters Xbridge C18 19×100 mm;gradient over 8 min from 30 to 100% of solvent B; solvent A: 10% MeOH:90% H₂O: 0.1% TFA; solvent B: 90% MeOH, 10% H₂O, 0.1% TFA),concentration, basification with aqueous K₂CO₃ solution, and extractionwith ethyl acetate gave((1R,3S)-1-amino-3-((S)-6-((phenylsulfonyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(2 mg, 4.76 μmol) as a solid. LC/MS M⁺¹=400. HPLC Retention time=7.04min (condition L) ¹H NMR (400 MHz, METHANOL-d₄) δ 7.99-7.93 (m, 2H),7.77-7.70 (m, 1H), 7.68-7.61 (m, 2H), 7.01-6.87 (m, 3H), 3.53-3.42 (m,2H), 3.27 (dd, J=6.3, 5.0 Hz, 2H), 3.07-2.88 (m, 2H), 2.80-2.71 (m, 2H),2.55 (dd, J=16.3, 9.9 Hz, 1H), 2.40-2.14 (m, 2H), 2.09-1.95 (m, 2H),1.95-1.67 (m, 3H), 1.64-1.49 (m, 2H).

The examples in Table 19 were prepared according to the generalprocedures for Examples 336 and 337.

TABLE 19 HPLC Ex. ret. time HPLC No. Structure MW (min.) condition MS(M⁺¹) 338

363.6 2.21 C 364 339

379.6 2.17 C 380

Example 343((1R,3S)-1-amino-3-((S)-6-hexyl-3-iodo-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol, TFA

To a solution of((1R,3S)-1-amino-3-((S)-6-hexyl-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (see PCT/US2014/017534) (10 mg, 0.023 mmol) in TFA (1 mL) was addedNIS (15.22 mg, 0.068 mmol) at room temperature. After 1 h, LCMS showedcomplete consumption of starting material. The solvent was removed andpurification on HPLC prep was performed. HPLC: condition=2 mL injection,gradient time of 5 min, start B=20% to 100%, stop time of 15 min,Solvent A=0.1% TFA in water, Solvent B=0.1% TFA in MeCN, column=LUNA,wavelength of 220 nm.((1R,3S)-1-amino-3-((S)-6-hexyl-3-iodo-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol, TFA (3.5 mg, 5.78 μmol) was isolated with >95% purity. HPLCretention time=12.3 min (condition L) LC/MS M⁺¹=456. ¹H NMR (400 MHz,METHANOL-d₄) δ 7.56 (s, 1H), 7.08 (s, 1H), 3.54-3.41 (m, 2H), 3.01 (tt,J=11.1, 7.2 Hz, 1H), 2.87-2.69 (m, 3H), 2.34 (dd, J=16.2, 10.5 Hz, 1H),2.20 (dd, J=13.0, 7.5 Hz, 1H), 2.07-1.84 (m, 3H), 1.83-1.60 (m, 3H),1.60-1.48 (m, 1H), 1.47-1.25 (m, 11H), 1.00-0.88 (m, 3H).

The examples in Table 20 were prepared according to the generalprocedure in Example 343.

TABLE 20 HPLC Ex. ret. time HPLC MS No. Structure MW (min.) method (M⁺¹)344

455.4 12.2 L 456 345

363.9 10.9 L 364/ 366

Example 346((1R,3S)-1-amino-3-((S)-6-hexyl-3-methyl-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol, TFA

To a solution of((1R,3S)-1-amino-3-((S)-6-hexyl-3-iodo-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (90 mg, 0.158 mmol) (dried with toluene evaporation) and ferricacetylacetonate (11.16 mg, 0.032 mmol) in a mixture of THF (1.5 mL) andN-methyl-2-pyrrolidinone (0.3 mL) was added methylmagnesium bromide(0.263 mL, 0.790 mmol) at room temperature. LCMS showed desired productalong with SM and desiodo product. The mixture was injected throughHPLC. HPLC: condition=2 mL injection, gradient time of 5 min, startB=20% to 100%, stop time of 15 min, Solvent A=0.1% TFA in Water, SolventB=0.1% TFA in MeCN, column=LUNA, wavelength of 220.((1R,3S)-1-amino-3-((S)-6-hexyl-3-methyl-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (15 mg, 0.031 mmol) was isolated with >95% purity. HPLC retentiontime=11.6 min (condition L) LC/MS M⁺¹=344. ¹H NMR (400 MHz, METHANOL-d₄)δ 7.05-6.99 (m, 1H), 7.08 (s, 1H), 3.54-3.41 (m, 2H), 3.01 (tt, J=11.1,7.2 Hz, 1H), 2.87-2.69 (m, 3H), 2.34 (dd, J=16.2, 10.5 Hz, 1H), 2.26 (s,3H), 2.20 (dd, J=13.0, 7.5 Hz, 1H), 2.07-1.84 (m, 3H), 1.83-1.60 (m,3H), 1.60-1.48 (m, 1H), 1.47-1.25 (m, 11H), 1.00-0.88 (m, 3H).

Example 3476-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-ylHexanoate, TFA

Preparation 347A:(5R,7S)-7-(6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of(5R,7S)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(200 mg, 0.701 mmol) in MeOH (7009 μl) at 0° C. was added sodiumborohydride (53.0 mg, 1.402 mmol) in one portion. The reaction mixturewas stirred at 0° C. for 30 min and then allowed to warm to roomtemperature. LCMS showed completion. The solvent was removed, the slurrywas diluted with DCM and washed twice with DCM. The organic layer wasdried with Na₂SO₄ and concentrated to afford(5R,7S)-7-(6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(201 mg, 0.699 mmol). The material was used directly for furtherreaction. HPLC retention time=0.75 min (condition G); LC/MS M⁺¹=288.

Preparation 347B:6-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-ol

To a mixture of(5R,7S)-7-(6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(200 mg, 0.696 mmol) in dioxane (5 mL) was added 1N NaOH (6.96 mL, 6.96mmol). The reaction mixture was heated at 100° C. for 14 h. LCMS showedcomplete consumption of starting material. The reaction mixture wasdiluted with ethyl acetate and washed with H₂O. The organic layer wasdried with MgSO₄, filtered and concentrated. The organic layer was driedwith MgSO₄, filtered, and concentrated to afford6-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-ol(180 mg, 0.689 mmol). HPLC retention time=4.9 min (condition L) LC/MSM⁺¹=262. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.06-6.95 (m, 3H), 4.10-3.96(m, 1H), 3.73-3.52 (m, 2H), 3.16-3.07 (m, 1H), 3.02 (dd, J=16.3, 4.6 Hz,1H), 2.98-2.87 (m, 1H), 2.82 (dd, J=9.5, 5.9 Hz, 1H), 2.68 (dd, J=16.2,8.0 Hz, 1H), 2.48-2.37 (m, 1H), 2.16-2.08 (m, 1H), 2.08-1.99 (m, 1H),1.99-1.86 (m, 3H), 1.83-1.67 (m, 2H).

Preparation 347C:((1R,3S)-3-(6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-1-(hydroxymethyl)cyclopentyl)carbamate

To a solution of6-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-ol(182 mg, 0.696 mmol) in DCM (6964 μl) was added BOC₂O (243 μl, 1.045mmol) and triethylamine (146 μl, 1.045 mmol). The reaction mixture wasstirred at room temperature overnight and LCMS showed completeconsumption of starting material. The reaction mixture was diluted withethyl acetate and washed with 1N HCl. The organic layer was dried withMgSO₄, filtered, and concentrated to afford tert-butyl((1R,3S)-3-(6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-1-(hydroxymethyl)cyclopentyl)carbamate (250 mg, 0.692 mmol) as an oil. HPLC retentiontime=0.89 min (condition G); LC/MS M⁺¹=364.

Preparation 347D: (5R,7S)-tert-butyl7-(6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-2,2-dimethyl-3-oxa-1-azaspiro[4.4]nonane-1-carboxylate

To a solution of tert-butyl((1R,3S)-3-(6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-1-(hydroxymethyl)cyclopentyl)carbamate(250 mg, 0.692 mmol) in acetone (6916 μl) was added 2,2-dimethoxypropane(170 μl, 1.383 mmol) followed by BF₃.OEt₂ (175 μl, 1.383 mmol). Thereaction was monitored by LCMS and after 1 h a considerable amount ofdesired product (RT of 1.14 min) was formed. The reaction was quenchedwith 0.5 mL Et₃N to complex BF₃. The solvent was removed under reducedpressure and placed under vacuum to afford (5R,7S)-tert-butyl7-(6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-2,2-dimethyl-3-oxa-1-azaspiro[4.4]nonane-1-carboxylate(278 mg, 0.692 mmol). HPLC retention time=1.16 min (condition G); LC/MSM⁺¹=402.

To a solution a solution of (5R,7S)-tert-butyl7-(6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-2,2-dimethyl-3-oxa-1-azaspiro[4.4]nonane-1-carboxylate(0.040 g, 0.1 mmol) in DCM (1.000 ml) was added pyridine (0.024 ml,0.300 mmol) and hexanoyl chloride (0.028 ml, 0.200 mmol) at roomtemperature. LCMS showed a rapid conversion to the desired product at1.41 min. HPLC retention time=1.41 min (condition G); LC/MS M⁺¹=500.4.To this solution was added TFA (1 mL) and the reaction was followed byLCMS. LCMS showed rapid conversion to the desired compound. The mixturewas concentrated under reduced pressure, dissolved in MeOH and purifiedby HPLC prep:HPLC:condition=2 mL injection, gradient time of 5 min,start B=20% to 100%, stop time of 15 min, Solvent A=0.1% TFA in Water,Solvent B=0.1% TFA in MeCN, column=LUNA, wavelength of 220.6-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-ylhexanoate, TFA (16 mg, 0.030 mmol) was isolated as a colorless oil withpurity >95%. HPLC retention time=0.89 min (condition G) LC/MS M⁺¹=361.¹H NMR (400 MHz, METHANOL-d₄) δ 7.11-6.92 (m, 3H), 5.24-5.13 (m, 1H),3.73-3.54 (m, 2H), 3.08 (dd, J=16.6, 4.7 Hz, 2H), 3.01-2.88 (m, 1H),2.88-2.77 (m, 2H), 2.50-2.38 (m, 1H), 2.36-2.25 (m, 2H), 2.13 (d, J=2.9Hz, 1H), 2.09-1.87 (m, 4H), 1.74 (t, J=12.8 Hz, 1H), 1.61 (quin, J=7.1Hz, 3H), 1.43-1.22 (m, 4H), 0.96-0.83 (m, 3H).

The examples in Table 21 were prepared according to the generalprocedures in Example 347.

TABLE 21 HPLC Ex. ret. time HPLC MS No. Strcture MW (min.) method (M⁺1)Comment 348

387.6 0.84 G 388 349

387.6 9.00 L 388 Isomer 1 350

387.6 9.05 L 388 Isomer 2 351

360.5 6.9 L 361 352

388.3 7.9 L 389 353

365.5 7.6 L 366Separation procedure for Examples 349 and 350: PreparativeChromatographic Conditions: Instrument: Berger SFC MGII; Column: ChiralAD-H 25×3 cm ID, 5 μm; Flow rate: 85.0 mL/min; Mobile Phase: 85/15CO₂/MeOH w/0.1% DEA; Detector 5 Wavelength: 220 nm; Sample Prep and Inj.Volume: 2500 μL of 20 mg dissolved in 6 mL MeOH/ACN. AnalyticalChromatographic Conditions: Instrument: Berger analytical SFC; Column:Chiral AD-H 250×4.6 mm ID, 5 μm; Flow rate: 2.0 mL/min; Mobile Phase:80/20 CO₂/MeOH w/0.1% DEA.

Example 3546-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-ylbutyl(methyl)carbamate, TFA

Preparation 354A: (5R,7S)-tert-butyl7-(6-((butyl(methyl)carbamoyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-2,2-dimethyl-3-oxa-1-azaspiro[4.4]nonane-1-carboxylate

To a solution of (5R,7S)-tert-butyl7-(6-((butylcarbamoyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-2,2-dimethyl-3-oxa-1-azaspiro[4.4]nonane-1-carboxylate(0.050 g, 0.1 mmol) in THF (2 mL) was added potassium tert-butoxide(0.045 g, 0.400 mmol) followed by Mel (0.025 mL, 0.400 mmol). Thereaction mixture was stirred 1 h at room temperature when LCMS showedcomplete consumption. The mixture was diluted with EtOAc and washedtwice with 1N HCl, dried over MgSO₄, filtered and concentrated underreduced pressure. The resulting material was used directly in the nextreaction. HPLC retention time=1.95 min (condition G); LC/MS M⁺¹=515.

Example 354

To a solution of (5R,7S)-tert-butyl7-(6-((butyl(methyl)carbamoyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-2,2-dimethyl-3-oxa-1-azaspiro[4.4]nonane-1-carboxylate(51.5 mg, 0.1 mmol) in DCM (2 mL) was added TFA (1 mL). The solution wasstirred for 30 min at room temperature when LCMS showed completeconsumption. The solvent was removed under reduced pressure and theresulting oil was dissolved in MeOH. The solution was injected on theHPLC. Condition=2 mL injection, gradient time of 5 min, start B=20% to100%, stop time of 15 min, Solvent A=0.1% TFA in water, Solvent B=0.1%TFA in MeCN, column=LUNA, wavelength of 220 providing6-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-ylbutyl(methyl)carbamate, TFA (12 mg, 0.023 mmol) with >95% purity. HPLCretention time=7.31 min (condition L); LC/MS M⁺¹=375. ¹H NMR (400 MHz,METHANOL-d₄) δ 7.12-6.97 (m, 3H), 5.15-4.99 (m, 1H), 3.64 (dd, J=13.9,12.5 Hz, 2H), 3.20-3.01 (m, 4H), 3.01-2.75 (m, 6H), 2.43 (dd, J=12.2,6.9 Hz, 1H), 2.20-2.09 (m, 1H), 2.09-1.89 (m, 5H), 1.73 (t, J=12.8 Hz,1H), 1.61-1.47 (m, 1H), 1.47-1.25 (m, 2H), 1.25-1.07 (m, 1H), 0.97 (t,J=7.2 Hz, 1.5H), 0.87-0.72 (m, 1.5H) mixture of 1:1 rotamer.

Example 355N-(6-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-yl)hexanamide,TFA

Preparation 355A:(5R,7S)-7-(6-amino-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of(5R,7S)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(50 mg, 0.175 mmol) in MeOH (1752 μl) was added ammonium acetate (135mg, 1.752 mmol) followed by sodium cyanoborohydride (16.52 mg, 0.263mmol). The reaction mixture was stirred at room temperature overnight.

LCMS showed complete consumption of starting material. Next, 4 mL of 1NHCl was added and the solvent was removed under reduced pressure. DCMwas added and the organic layer was washed twice with 1N HCl. Theaqueous layer was basified with 1N NaOH and extracted 3 times withEtOAc. The organic fractions were combined, dried and concentratedaffording(5R,7S)-7-(6-amino-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(44 mg, 0.154 mmol) as a brown oil. The material was used directly forfurther reaction. HPLC retention time=0.55 min (condition G); LC/MSM+=287.

Preparation 355B:((1R,3S)-1-amino-3-(6-amino-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a solution of(5R,7S)-7-(6-amino-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(44 mg, 0.154 mmol) in dioxane (1536 μl) in a vial was added NaOH (1536μl, 1.536 mmol). The vial was sealed and warmed to 100° C. for 1 h. LCMSshowed complete conversion. Next, 4 mL NaOH 1N was added and the aqueouslayer was washed three times with EtOAc. The organics layers werecombined, dried and concentrated affording((1R,3S)-1-amino-3-(6-amino-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol (20 mg, 0.077 mmol) as an oil. HPLC retentiontime=0.42 min (condition G); LC/MS M⁺¹=261.

Example 355

To a solution of((1R,3S)-1-amino-3-(6-amino-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(10 mg, 0.038 mmol) in DCM (384 μl) was added hexanoyl chloride (6.44μl, 0.046 mmol). The reaction mixture was stirred at 25° C. for 15 minand the reaction was quenched with 1N NaOH. The aqueous layer wasextracted 3 times with EtOAc. The organic layers were combined, driedand concentrated under reduced pressure. The resulting oil was dissolvedin MeOH. The solution was injected on the HPLC prep: condition=2 mLinjection, gradient time of 5 min, start B=20% to 100%, stop time of 15min, Solvent A=0.1% TFA in water, Solvent B=0.1% TFA in MeCN,column=LUNA, wavelength of 220 nm.N-(6-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-yl)hexanamide,TFA (3.7 mg, 7.44 μmol) was obtained with >95% purity. HPLC retentiontime=6.59 min (condition L) LC/MS M⁺¹=359. ¹H NMR (400 MHz, METHANOL-d₄)δ 7.10-6.98 (m, 3H), 4.13-4.00 (m, 1H), 3.64 (dd, J=13.6, 11.0 Hz, 2H),3.20-3.07 (m, 1H), 3.01 (dd, J=16.0, 4.5 Hz, 1H), 2.90 (dd, J=8.0, 5.0Hz, 2H), 2.65 (dd, J=16.3, 9.7 Hz, 1H), 2.43 (dd, J=13.3, 6.1 Hz, 1H),2.21 (t, J=7.5 Hz, 2H), 2.17-2.01 (m, 2H), 2.01-1.87 (m, 3H), 1.79-1.57(m, 4H), 1.44-1.27 (m, 4H), 0.94 (t, J=7.0 Hz, 3H).

Example 356 in Table 22 was prepared according to the general procedurefor Example 355.

TABLE 22 HPLC Ex. ret. time HPLC MS No. Structure MW (min.) method (M⁺¹)356

359.5 0.61 G 360

Example 357((1R,3S)-1-amino-3-(6′-butyl-3,3′,4,4′,5′,6′-hexahydro-1H-spiro[naphthalene-2,2′-pyran]-6-yl)cyclopentyl)methanol

Preparation 357A:5R,7S)-7-(6′-butyl-4′-chloro-3,3′,4,4′,5′,6′-hexahydro-1H-spiro[naphthalene-2,2′-pyran]-6-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of(5R,7S)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(200 mg, 0.701 mmol) and oct-1-en-4-ol (180 μl, 1.402 mmol) in CH₂Cl₂(7009 μl) at room temperature was added tin(IV) chloride (841 μl, 0.841mmol). LCMS showed that the reaction was complete within 2 h. Thereaction was quenched by saturated NaHCO₃ and the aqueous layer was backextract three times with DCM. The organic layers were combined, driedand concentrated under reduced pressure. The resulting oil was purifiedby HPLC prep (condition=2 mL injection, gradient time of 5 min, startB=20% to 100%, stop time of 15 min, Solvent A=0.1% TFA in Water, SolventB=0.1% TFA in MeCN, column=LUNA, wavelength of 220) affording(5R,7S)-7-(6′-butyl-4′-chloro-3,3′,4,4′,5′,6′-hexahydro-1H-spiro[naphthalene-2,2′-pyran]-6-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(149 mg, 0.345 mmol) as a brown solid which was used directly forfurther reaction. HPLC retention time=1.18 min (condition G) LC/MSM⁺¹=432/434.

Preparation 357B:(5R,7S)-7-(6′-butyl-3,3′,4,4′,5′,6′-hexahydro-1H-spiro[naphthalene-2,2′-pyran]-6-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of(5R,7S)-7-(6′-butyl-4′-chloro-3,3′,4,4′,5′,6′-hexahydro-1H-spiro[naphthalene-2,2′-pyran]-6-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(300 mg, 0.694 mmol) in i-PrOH (3472 μl) was added HCl (6945 μl, 41.7mmol) followed by zinc (4540 mg, 69.4 mmol). The reaction mixture waswarmed to 80° C. and followed by LCMS.

Conversion of >90% after 3 days was measured. The heterogeneous mixturewas filtered through Celite eluting with EtOAc. The oil obtained afterconcentration under reduced pressure was purified by HPLC prep(condition=2 mL injection, gradient time of 5 min, start B=20% to 100%,stop time of 15 min, Solvent A=0.1% TFA in Water, Solvent B=0.1% TFA inMeCN, column=LUNA, wavelength of 220) affording(5R,7S)-7-(6′-butyl-3,3′,4,4′,5′,6′-hexahydro-1H-spiro[naphthalene-2,2′-pyran]-6-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(100 mg, 0.252 mmol) as a white solid which was used directly forfurther reaction. HPLC retention time=1.21 min (condition G) LC/MSM⁺¹=398.3. Stereoisomer separation was then performed. Approximately 110mg sample was resolved. Four isomers were collected. PreparativeChromatographic Conditions: Instrument: Berger SFC MGII; Column: ChiralOJ-H 25×3 cm ID, 5 μm; Flow rate:

85.0 mL/min; Mobile Phase: 85/15 C02/1:1 MeOH:CAN; Detector Wavelength:220 nm; Sample Prep and Inj. Volume: 1000 μL of 1100 mg dissolved in 7mL MeOH/ACN. Analytical Chromatographic Conditions: Instrument: Bergeranalytical SFC; Column: Chiral OJ-H 250×4.6 mm ID, 5 μm; Flow rate: 2.0mL/min; Mobile Phase: 87/13 C02/1:1 MeOH:CAN.

Example 357:((1R,3S)-1-amino-3-(6′-butyl-3,3′,4,4′,5′,6′-hexahydro-1H-spiro[naphthalene-2,2′-pyran]-6-yl)cyclopentyl)methanol,TFA (Isomer 1)

To a solution of Isomer I of(5R,7S)-7-(6′-butyl-3,3′,4,4′,5′,6′-hexahydro-1H-spiro[naphthalene-2,2′-pyran]-6-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(17 mg, 0.043 mmol) in dioxane (428 μl) was added NaOH (428 μl, 0.428mmol). The reaction mixture was warmed to 100° C. LCMS showed completeconversion after 2 h. The reaction mixture was cooled down to roomtemperature, diluted with EtOAc and 1N NaOH. The aqueous layer wasextracted three times with EtOAc. The organic layers were combined,dried, and concentrated under reduced pressure. The resulting oil wasdiluted in MeOH and injected on HPLC: condition=2 mL injection, gradienttime of 5 min, start B=20% to 100%, stop time of 15 min, Solvent A=0.1%TFA in Water, Solvent B=0.1% TFA in MeCN, column=LUNA, wavelength of 220affording((1R,3S)-1-amino-3-(6′-butyl-3,3′,4,4′,5′,6′-hexahydro-1H-spiro[naphthalene-2,2′-pyran]-6-yl)cyclopentyl)methanol,TFA isomer 1 (15 mg, 0.031 mmol). HPLC retention time=9.09 min(condition L). LC/MS M⁺¹=372. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.08-6.93(m, 3H), 3.72-3.52 (m, 3H), 3.21-3.04 (m, 1H), 2.92-2.75 (m, 1H),2.75-2.60 (m, 3H), 2.55-2.37 (m, 2H), 2.20-2.03 (m, 1H), 2.02-1.90 (m,3H), 1.90-1.78 (m, 1H), 1.73 (t, J=12.8 Hz, 2H), 1.68-1.45 (m, 4H),1.44-1.32 (m, 2H), 1.32-1.09 (m, 5H), 0.85 (t, J=7.0 Hz, 3H).

The examples in Table 23 were prepared according to the generalprocedure for Example 357

TABLE 23 HPLC Ex. ret. time HPLC No. Structure MW (min.) method MS (M⁺¹)Comment 358 359 360

371.6 371.6 371.6 8.96 8.98 8.96 L L L 372 372 372 Isomer 2 Isomer 3Isomer 4 361

315.5 6.68 L 316 —

Examples 362 to 3636-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-2-butyl-1,2,3,4-tetrahydronaphthalen-1-ol,TFA

Preparation 362A:(5R,7S)-7-(6-butyl-5-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of(5R,7S)-7-(6-butyl-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(0.5 g, 1.536 mmol) in THF (15.36 ml) was added BH₃.DMS (0.307 ml, 1.536mmol) at room temperature. The reaction mixture was stirred for 1.5 h.LCMS showed no starting material. To the reaction mixture was added 1NNaOH (0.5 mL) and H₂O₂ (1.569 ml, 15.36 mmol) and the reaction mixturewas stirred at room temperature for 15 min. The reaction mixture wasdiluted with water, extracted with EtOAc, and then washed with water(2×). The organic layers were combined, dried over Na₂SO₄, andconcentrated under reduced pressure. The resulting oil was purified onISCO affording(5R,7S)-7-(6-butyl-5-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(0.34 g, 0.990 mmol). ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.45 (d, J=7.9Hz, 1H), 7.07 (d, J=7.9 Hz, 1H), 6.95 (s, 1H), 6.22-6.08 (m, 1H), 4.41(d, J=6.6 Hz, 1H), 4.37-4.23 (m, 2H), 3.11-2.92 (m, 2H), 2.84-2.69 (m,2H), 2.37-2.24 (m, 1H), 2.21-2.02 (m, 3H), 2.02-1.88 (m, 2H), 1.88-1.60(m, 4H), 1.57-1.51 (m, 1H), 1.50-1.44 (m, 1H), 1.42-1.25 (m, 4H),1.03-0.86 (m, 3H).

Examples 362 and 363

To a solution of(5R,7S)-7-(6-butyl-5-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(22 mg, 0.064 mmol) in dioxane (320 μl) was added NaOH (641 μl, 0.641mmol). The reaction mixture was warmed to 90° C. and stirred until fullconversion was observed (2 h). The reaction mixture was cooled, dilutedwith water and EtOAc. The aqueous layer was back extract three timeswith EtOAc. The organic layers were combined, dried and concentratedunder reduced pressure. The resulting oil was solubilized in MeOH andpurified by HPLC: condition=2 mL injection, gradient time of 5 min,start B=20% to 100%, stop time of 15 min, Solvent A=0.1% TFA in Water,Solvent B=0.1% TFA in MeCN, column=LUNA, wavelength of 220 nm.6-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-2-butyl-1,2,3,4-tetrahydronaphthalen-1-ol,TFA (9 mg, 0.02 mmol) was obtained with >95% purity. PreparativeChromatographic Conditions: Instrument: Berger SFC MGII; Column: ChiralOD-H 25×3 cm ID, 5 μm; Flow rate: 85.0 mL/min; Mobile Phase: 75/25CO₂/MeOH; Detector Wavelength: 220 nm; Sample Preparation and Inj.Volume: 700 μL-1000 μL of 52 mg dissolved in 2 mL MeOH. AnalyticalChromatographic Conditions: Instrument: Berger analytical SFC; Column:Chiral OD-H 250×4.6 mm ID, 5 μm; Flow rate: 2.0 mL/min; Mobile Phase:80/20 CO₂/MeOH.

Example 362: Isomer 1: HPLC retention time=0.77 min (condition G); LC/MSM+=318; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.41 (d, J=8.1 Hz, 1H),7.17-7.07 (m, 1H), 7.01 (s, 1H), 4.34 (d, J=6.8 Hz, 1H), 3.64 (dd,J=15.6, 11.7 Hz, 2H), 3.21-3.06 (m, 1H), 2.77 (t, J=6.3 Hz, 2H), 2.43(dd, J=13.4, 7.0 Hz, 1H), 2.18-2.04 (m, 2H), 2.03-1.88 (m, 3H),1.81-1.63 (m, 3H), 1.59-1.44 (m, 2H), 1.44-1.30 (m, 3H), 1.30-1.17 (m,1H), 1.02-0.90 (m, 3H).

Example 363: Isomer 2: HPLC retention time=0.78 min (condition G); LC/MSM⁺¹=318; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.41 (d, J=8.1 Hz, 1H),7.17-7.07 (m, 1H), 7.01 (s, 1H), 4.34 (d, J=6.8 Hz, 1H), 3.64 (dd,J=15.6, 11.7 Hz, 2H), 3.21-3.06 (m, 1H), 2.77 (t, J=6.3 Hz, 2H), 2.43(dd, J=13.4, 7.0 Hz, 1H), 2.18-2.04 (m, 2H), 2.03-1.88 (m, 3H),1.81-1.63 (m, 3H), 1.59-1.44 (m, 2H), 1.44-1.30 (m, 3H), 1.30-1.17 (m,1H), 1.02-0.90 (m, 3H).

Examples 364 to 366(5R,7S)-7-(6-butyl-5-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of(5R,7S)-7-(6-butyl-5-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(150 mg, 0.437 mmol) in DCM (2184 μl) was added DMP (370 mg, 0.873 mmol)at room temperature. LCMS showed complete conversion after 1 h. Thereaction mixture was diluted with DCM and extracted with 1N Na₂S₂O₃ and1N NaOH affording an oily compound after concentration under reducedpressure. The resulting oil was solubilized in MeOH and purified byHPLC: condition=2 mL injection, gradient time of 5 min, start B=20% to100%, stop time of 15 min, Solvent A=0.1% TFA in Water, Solvent B=0.1%TFA in MeCN, column=LUNA, wavelength of 220.(5R,7S)-7-(6-butyl-5-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(128 mg, 0.375 mmol) was obtained with >95% purity. HPLC retentiontime=1.24 min (condition G); LC/MS M⁺¹=342; ¹H NMR (400 MHz,CHLOROFORM-d) δ 7.98 (d, J=8.1 Hz, 1H), 7.16 (dd, J=8.1, 1.5 Hz, 1H),7.10 (s, 1H), 6.59 (s, 1H), 4.33 (dd, J=13.2, 8.1 Hz, 2H), 3.15-3.02 (m,1H), 3.02-2.91 (m, 2H), 2.52-2.41 (m, 1H), 2.35 (dd, J=13.2, 7.3 Hz,1H), 2.31-2.09 (m, 3H), 2.07-1.78 (m, 6H), 1.60-1.45 (m, 1H), 1.45-1.31(m, 4H), 1.02-0.86 (m, 3H). Preparative Chromatographic Conditions:Instrument: Berger SFC MGII; Column: Chiral OD-H 25×3 cm ID, 5 μm; Flowrate: 80.0 mL/min; Mobile Phase: 75/25 CO₂/MeOH; Detector Wavelength:220 nm; Sample Prep and Inj. Volume: 700 μL-1000 μL of 52 mg dissolvedin 2 mL MeOH. Analytical Chromatographic Conditions: Instrument: Bergeranalytical SFC; Column: Chiral OD-H 250×4.6 mm ID, 5 μm; Flow rate: 2.0mL/min; Mobile Phase: 75/25 CO₂/MeOH.

Example 365: Isomer 1: HPLC retention time=1.24 min (condition G); LC/MS

M⁺¹=342; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.89 (d, J=8.1 Hz, 1H),7.35-7.16 (m, 2H), 3.60-3.46 (m, 2H), 3.17 (ddd, J=11.2, 7.0, 3.7 Hz,1H), 3.10-2.92 (m, 2H), 2.57-2.43 (m, 1H), 2.39-2.18 (m, 2H), 2.17-2.04(m, 1H), 2.04-1.75 (m, 6H), 1.63 (t, J=12.3 Hz, 1H), 1.58-1.34 (m, 4H),1.01-0.89 (m, 3H).

Example 366: Isomer 2: HPLC retention time=1.24 min (condition G); LC/MS

M⁺¹=342; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.89 (d, J=8.1 Hz, 1H),7.35-7.16 (m, 2H), 3.60-3.46 (m, 2H), 3.17 (ddd, J=11.2, 7.0, 3.7 Hz,1H), 3.10-2.92 (m, 2H), 2.57-2.43 (m, 1H), 2.39-2.18 (m, 2H), 2.17-2.04(m, 1H), 2.04-1.75 (m, 6H), 1.63 (t, J=12.3 Hz, 1H), 1.58-1.34 (m, 4H),1.01-0.89 (m, 3H).

Example 367((1R,3S)-1-amino-3-((R)-6-((E)-hex-1-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 367A: 5-(pentylsulfonyl)-1-phenyl-1H-tetrazole

DEAD (727 μl, 4.59 mmol) was added dropwise to a solution of pentan-1-ol(300 mg, 3.40 mmol), 1-phenyl-1H-tetrazole-5-thiol (740 mg, 4.15 mmol)and Ph₃P (1089 mg, 4.15 mmol) in THF (20 ml) at 0° C. The mixture wasstirred at a temperature range of from 0° C. to room temperature for 16h. The mixture was diluted with EtOAc (30 ml), which was washed withbrine (2×20 ml), water (20 ml) and brine (20 ml), dried (Na₂SO₄) andconcentrated under vacuo. The residue was purified with flashchromatography using Isco column (25 g, EtOAc/Hexane=0-50%, gradienttime=25 min) to get 5-(pentylthio)-1-phenyl-1H-tetrazole (650 mg). LC/MSM⁺¹=249. Ammonium molybdate tetrahydrate (679 mg, 0.550 mmol) was addedin 30% H₂O₂ (4064 μl, 39.8 mmol) at 0° C. and the resultant solution wasadded dropwise to a solution of 5-(pentylthio)-1-phenyl-1H-tetrazole(650 mg, 2.62 mmol) in EtOH (20 ml) at 0° C., the mixture was allowed towarm to room temperature and stirred at room temperature for 16 h. Next,30 ml of brine was added and the mixture was extracted with EtOAc (80ml), which was washed with brine and dried (Na₂SO₄), concentrated undervacuo to give the desired product which was used as is.5-(pentylsulfonyl)-1-phenyl-1H-tetrazole (700 mg). LC/MS M⁺¹=281.

Preparation 367B:(5R,7S)-7-((R)-6-((E)-hex-1-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

KHMDS (418 μl, 0.209 mmol) was added dropwise to a solution of5-(pentylsulfonyl)-1-phenyl-1H-tetrazole (25.8 mg, 0.092 mmol) and(R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carbaldehyde(25 mg, 0.084 mmol) in THF (3 ml) at −78° C. and the resultant solutionwas stirred at the temperature for 1 h. H₂O (1 ml) was added and themixture was warmed to room temperature and 30 ml of brine was added andthe mixture was extracted with EtOAc (80 ml), which was washed withbrine and dried (Na₂SO₄), concentrated under vacuo to afford(5R,7S)-7-((R)-6-((E)-hex-1-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(5 mg). LC/MS M⁺¹=354.

Example 367

(5R,7S)-7-((R)-6-((E)-hex-1-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(5 mg, 0.014 mmol) in 1,4-dioxane (2 ml) was mixed with water (0.5 ml)and lithium hydroxide hydrate (5.94 mg, 0.141 mmol) was added. Themixture was stirred at 100° C. for 16 h under N₂. After cooling, themixture was filtered and washed with MeOH, the combined solvents wereevaporated and the residue was purified with prep HPLC: columnPhenomenex Luna C18 5u 21.2×100 mm. Solvent A: 10% MeOH-90% H₂O-0.1%TFA; Solvent B: 90% MeOH-10% H₂O-0.1% TFA. Gradient time=15 min. StartB=0%, Final B 100%. Stop time 20 min. The collected fraction wasbasified with saturated NaHCO₃, concentrated under vacuo and the aqueouslayer was extracted with DCM (3×20 ml), which was dried (Na₂SO₄) andconcentrated under vacuo. The residue was freeze dried to afford((1R,3S)-1-amino-3-((R)-6-((E)-hex-1-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(3 mg). LC/MS M⁺¹=328. HPLC Rt=8.52 (condition L). ¹H NMR (400 MHz,METHANOL-d₄) δ 7.05-6.88 (m, 3H), 5.59-5.30 (m, 2H), 3.60-3.45 (m, 2H),3.10-2.98 (m, 1H), 2.89-2.74 (m, 3H), 2.64-2.18 (m, 3H), 2.11-1.74 (m,8H), 1.69-1.46 (m, 3H), 1.02-0.85 (m, 5H).

The examples in Table 24 were prepared according to the generalprocedure of Example 367.

TABLE 24 HPLC Ex. ret. time HPLC MS No. Structure MW (min.) method (M⁺¹)368

327.5 8.52 L 328 369

343.5 6.87 L 344 370

325.5 7.63 L 326 371

343.5 6.76 L 344 372

377.5 7.63 L 378 373

355.5 6.52 L 356 374

377.5 7.54 L 378 375

361.5 8.13 L 362 376

341.5 9.05 L 342

The following olefins were made according to the method listed in thetable.

TABLE 25 HPLC Ex. ret. time HPLC MS No. Structure MW (min.) method (M⁺¹)Method 377

392.5 8.23 L 330 See method for Example 226 378

325.5 8.26 L 326 See method for Example 226 379

327.5 8.37 L 328 See method for alternative Preparation- 2 of Example679 380

327.5 8.75 L 328 See method for alternative Preparation- 2 of Example679

Example 381((1R,3S)-1-amino-3-((R)-6-((E)-hex-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 381A:(5R,7S)-7-((R)-6-(hex-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

A mixture of trans-3-hexene (5.8 mL, 46.7 mmol),(5R,7S)-7-((R)-6-(but-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(1.5 g, 4.61 mmol) and dichloromethane (50 mL) was bubbled with nitrogenfor 3 min at −78° C. before Grubbs catalyst 2nd generation (0.25 g,0.294 mmol) was added. The bubbling was continued for 2 min. The mixturewas then stirred under nitrogen at 40° C. for 3.5 h. The mixture wasconcentrated. Flash chromatography purification (24 g silica gel column,gradient elution from 0 to 40% of ethyl acetate in DCM) afforded(5R,7S)-7-((R)-6-(hex-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(1.2 g, 3.39 mmol) as a solid. SFC separation (20% MeOH in C02, ADHcolumn; 40° C.; 140 bar BPR) gave PK1:(5R,7S)-7-((R)-6-((E)-hex-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(0.8 g, 2.263 mmol) (HPLC retention time=4.11 min (condition C); LC/MSM⁺¹=354); and PK2:(5R,7S)-7-((R)-6-((Z)-hex-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(0.1 g, 0.283 mmol) (HPLC retention time=4.08 min (condition C); LC/MSM⁺¹=354) as solids.

Example 381

A mixture of(5R,7S)-7-((R)-6-((E)-hex-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(0.41 g, 1.160 mmol), dioxane (5 mL), NaOH (0.928 g, 23.20 mmol), andwater (7 mL) was stirred at 90° C. under nitrogen for 1.5 days. Themixture was extracted with ethyl acetate (4×4 mL). The combined ethylacetate extracts were dried (Na₂SO₄) and concentrated. Purificationusing reverse phase HPLC (Phenomenex Luna 5μ 30×100 mm (Axia); gradientover 6 min from 40 to 100% of solvent B and holding @100% of solvent Bfor 6 min; solvent A: 10% MeOH: 90% H₂O: 0.10% TFA; solvent B: 90% MeOH,10% H₂O, 0.10% TFA), concentration, basification with 2N aqueous NaOH,and extraction with ethyl acetate gave((1R,3S)-1-amino-3-((R)-6-((E)-hex-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(0.34 g, 1.027 mmol) as a solid. PLC retention time=3.65 mi (conditionC); LC/MS M⁺¹=328. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.05-6.93 (m, 3H),5.56-5.36 (m, 2H), 3.53-3.40 (m, 2H), 3.09-2.96 (m, 1H), 2.88-2.74 (m,3H), 2.38 (dd, J=16.4, 10.5 Hz, 1H), 2.28 (dd, J=13.3, 8.0 Hz, 1H),2.15-1.85 (m, 8H), 1.83-1.62 (m, 2H), 1.52 (dd, J=13.2, 11.0 Hz, 1H),1.46-1.32 (m, 3H), 0.97 (t, J=7.5 Hz, 3H).

The examples in Table 26 were prepared according to the generalprocedure of Example 381.

TABLE 26 Ex. HPLC HPLC MS No. Structure MW ret. time (min.) method (M⁺¹)382

327.5 3.51 C 328 383

327.5 3.51 C 328 384

327.5 8.42 L 328 385

327.5 8.52 L 328 386

327.5 9.51 L 328 387

327.5 3.61 C 328 388

327.5 3.61 C 328 389

327.5 3.64 C 328 390

327.5 3.54 C 328 391

327.5 10.11 L 328 392

341.5 3.76 C 342 393

341.5 3.73 C 342 394

341.5 3.71 C 342 395

341.5 3.66 C 342 396

325.5 7.81 O 326 397

325.5 7.81 L 326

Examples 398 to 400(E)-6-(3-amino-3-(hydroxymethyl)cyclopentyl)-3,4-dihydronaphthalen-1(2H)-oneO-phenethyl oxime

Preparation 398A: Ethyl1-((diphenylmethylene)amino)-4-(5-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopent-2-enecarboxylate

A mixture was prepared by combining6-bromo-3,4-dihydronaphthalen-1(2H)-one (200 mg, 0.889 mmol), ethyl1-((diphenylmethylene)amino)cyclopent-3-enecarboxylate (426 mg, 1.333mmol), Et₃N (0.248 mL, 1.777 mmol) and 1,1′-bis(di-tert-butylphosphino)ferrocene palladium dichloride (29.0 mg, 0.044 mmol) into DL-tocopherolmethoxypolyethylene glycol succinate solution (2 wt % in H₂O) undernitrogen in a reaction vial. The vial was sealed and heated to 50° C.for 24 hours. The resulting mixture was poured into 200 ml ethylacetate. The solution was washed with water. The organic layer was thenconcentrated and the crude materials were purified on a 24 g silicacolumn (0-30% gradient ethyl acetate in hexane) to afford 260 mg oftitled compound. LC-MS Ret. Time: 1.69. LC-MS M⁺¹=464.2. LC-MSConditions: Column:Luna C18 4.6×30 mm 3u A:10:90 H₂O:ACN NH₄OAc/B:10:90H₂O:ACN NH₄OAc; 0%-95% B in 2 min; 4 mL/min flow. Product detected at220 nm wavelength.

Preparation 398B: Ethyl1-amino-3-(5-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentanecarboxylate

To a solution of ethyl1-((diphenylmethylene)amino)-4-(5-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopent-2-enecarboxylate(290 mg, 0.626 mmol) in EtOH (3 mL) was added 1.5 ml of 4N HCl. Thereaction mixture was allowed to stir at room temperature for 2 hours.LC-MS showed completed conversion. The mixture was poured into 50 ml ofsaturated NaHCO₃ and extracted with ethyl acetate twice. The organiclayer were then dried over Na₂SO₄ and concentrated to provide 270 mg ofcrude materials as yellow oil. The above obtained material was dissolvedinto ethyl acetate. To the solution was added Pd/C (107 mg, 0.050 mmol)under N₂. The mixture was allowed to stir under H₂ for 1 hour. LC-MSshowed the reaction was completed. The Pd catalyst was removed byfiltration. Solvent was removed to provide 260 mg of material. Thematerial was purified on a 40 g silica column (0%-30% gradient ethylacetate in hexane in 10 mins) to afford 170 mg of titled compound ascolorless oil. HPLC retention time=2.19 min (Condition K); LC/MSM⁺¹=302.

Preparation 398C: (E)-ethyl1-amino-3-(5-(phenethoxyimino)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentanecarboxylate

Ethyl 1-amino-3-(5-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentanecarboxylate (160 mg, 0.531 mmol) was dissolved into ethanol(4 mL). Pyridine (0.129 mL, 1.593 mmol) was added followed by theaddition of 0-phenethylhydroxylamine (124 mg, 0.903 mmol). The mixturewas allowed to stir at room temperature for 1 hour. LC-MS showed noreaction. The temperature was raised to 74° C. and the mixture wasallowed to stir for 9 hours. LC-MS showed <10% conversion. Thetemperature was raised to 85° C. and allowed to stir for 18 hours. LC-MSshowed 50% conversion. The heating was continued at 85° C. for 40 morehours. LC-MS showed >90% conversion. The reaction mixture was cooleddown to room temperature and poured into 50 ml saturated NaHCO₃. It wasextracted with ethyl acetate twice. The organic layers were then driedover Na₂SO₄ and concentrated to provide 290 mg of material having a HPLCpurity of 75%. HPLC retention time=0.97 min (Condition G); LC/MSM⁺¹=421.

Examples 398 to 400

(E)-ethyl1-amino-3-(5-(phenethoxyimino)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentanecarboxylate(290 mg, 0.690 mmol) was dissolved into MeOH (6 mL). The mixture wascooled to 0° C. and NaBH₄ (117 mg, 3.10 mmol) was added in portions. Themixture was allowed to stir for 3 hours. LC-MS showed partialconversion. The reaction mixture was allowed to stir at room temperaturefor 18 more hours. LC-MS showed completed conversion. The reaction wasquenched with 3 N HCl (aq.). The mixture was allowed to stir at roomtemperature for 30 mins. The crude material was purified on reversephase HPLC to provide 155 mg of product as white solid(diastereoisomeric mixture). HPLC retention time=0.94 min (Condition G);LC/MS M⁺¹=379.

Chiral SFC Separation of isomers: Approximately 100 mg sample wasresolved. The fractions (“Peak-1”, “Peak-2”, “Peak-3”, and “Peak-4”)were collected in MeOH w/0.1% DEA. The isomeric purity of each fractionwas estimated to be greater than 95% based on the prep-SFCchromatograms. Experimental Details: Preparative ChromatographicConditions: Instrument: Berger SFC MGII; Column: Phenomenex LUXCellulose 2 25×3 cm ID, 5 μm; Flow rate: 85.0 mL/min; Mobile Phase:65/35 CO₂/MeOH w/0.1% DEA; Detector Wavelength: 280 nm; Sample Prep andInj. Volume: 500 μL of 100 mg dissolved in 4.5 mL MeOH. AnalyticalChromatographic Conditions: Instrument: Berger analytical SFC; Column:Phenomenex LUX Cellulose 2 250×4.6 mm ID, 5 μm; Flow rate: 2.0 mL/min;Mobile Phase: 65/35 CO₂/MeOH w/0.1% DEA; Three fractions were isolatedin a 1:10:10 ratio having one minor peak and two major peaks. Example398: minor fraction, analytical SFC Retention Time: 12.58 min. Example399: major fraction 1, analytical SFC Retention Time: 13.87 min; HPLCretention time=0.88 mins (Condition G) LC/MS M⁺¹=379. ¹H NMR (400 MHz,METHANOL-d₄) δ 7.84 (d, J=8.1 Hz, 1H), 7.31-7.22 (m, 4H), 7.21-7.15 (m,1H), 7.15-7.06 (m, 2H), 4.34 (t, J=6.8 Hz, 2H), 3.54-3.42 (m, 2H),3.12-3.04 (m, 1H), 3.01 (t, J=6.8 Hz, 2H), 2.76-2.69 (m, 2H), 2.66 (t,J=6.6 Hz, 2H), 2.23 (dd, J=13.2, 7.7 Hz, 1H), 2.07-1.88 (m, 2H),1.85-1.66 (m, 4H), 1.60-1.51 (m, 1H). Example 400: major Fraction 2,analytical SFC Retention Time: 15.56 min; HPLC retention time=0.88 mins(Condition G) LC/MS M⁺¹=379. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.83 (d,J=8.1 Hz, 1H), 7.30-7.22 (m, 4H), 7.20-7.15 (m, 1H), 7.13-7.03 (m, 2H),4.33 (t, J=6.9 Hz, 2H), 3.53-3.42 (m, 2H), 3.10-3.03 (m, 1H), 3.02-2.97(m, 2H), 2.71 (t, J=6.1 Hz, 2H), 2.65 (t, J=6.7 Hz, 2H), 2.23 (dd,J=13.2, 7.5 Hz, 1H), 2.05-1.87 (m, 2H), 1.85-1.66 (m, 4H), 1.55 (t,J=12.3 Hz, 1H).

Example 401(E)-1-((R)-6-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-yl)ethanoneO-(2-methoxybenzyl) oxime

Preparation 401A:1-((R)-6-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-yl)ethanone

A solution of(5R,7S)-7-((R)-6-acetyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(200 mg, 0.638 mmol) in DCM (5 ml) and lithium hydroxide hydrate (402mg, 9.57 mmol) was mixed with THF (4 ml) and water (1 ml) and stirred at100° C. for overnight. The mixture was purified with prep HPLC:Phenomenex Luna C 18 5u (21.2×150 mm), Solvent A: 10% MeOH-90% H₂O-0.1%TFA; Solvent B: 90% MeOH-10% H₂O-0.1% TFA, Start B %=0, Final % B=100.Gradient time 15 min, stop time 22 min. (140 mg), LC/MS M⁺¹=288

Example 401

To a mixture of1-((S)-6-((1S,3R)-3-amino-3-(hydroxymethyl)cyclopentyl)-1,2,3,4-tetrahydronaphthalen-2-yl)ethanone(15 mg, 0.052 mmol) and O-(2-methoxybenzyl)hydroxylamine (40.0 mg, 0.261mmol) in EtOH (1.5 ml) was added 2 drops of 1N HCl. The mixture wasstirred at room temperature for 2 h. LC-MS indicated the completion ofconversion. The mixture was purified with prep HPLC: Phenomenex Luna C18 5u (21.2×150 mm), Solvent A: 10% MeOH-90% H₂O-0.1% TFA; Solvent B:90% MeOH-10% H₂O-0.1% TFA, Start B %=0, Final % B=100. Gradient time 15min, stop time 25 min. (15 mg as TFA salt). LC/MS M⁺¹=423. HPLC: Rt=7.50min (condition L). ¹H-NMR (400 MHz, METHANOL-d₄) δ 7.36-7.19 (m, 2H),7.10-6.86 (m, 5H), 5.18-5.05 (m, 2H), 3.90-3.79 (m, 3H), 3.72-3.56 (m,2H), 3.20-3.04 (m, 1H), 2.93-2.75 (m, 4H), 2.64-2.54 (m, 1H), 2.43 (dd,J=13.9, 6.6 Hz, 1H), 2.21-1.83 (m, 8H), 1.80-1.63 (m, 2H).

The examples in Table 27 were prepared according to the generalprocedure of Example 401.

TABLE 27 HPLC Ex. ret. time HPLC No. Structure MW (min.) method MS (M⁺¹)402

392.5 7.42 L 393 403

358.5 7.53 L 359 404

406.6 7.72 L 407 405

422.6 7.39 L 423 406

422.6 7.31 L 423 407

410.5 7.54 L 411 408

330.4 7.51 L 331 409

330.5 7.49 L 331 410

392.5 8.76 L 393 411

378.5 8.06 L 379

PHOSPHORYLATED EXAMPLES Example 412((1R,3S)-1-amino-3-((R)-2-((pentyloxy)methyl)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)cyclopentyl)methylDihydrogen Phosphate

To a stirred solution of((1R,3S)-1-amino-3-((R)-2-((pentyloxy)methyl)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)cyclopentyl)methanol(2.6 mg, 7.44 μmol) in anhydrous acetonitrile (1 mL) was addedpyrophosphoryl chloride (0.011 mL, 0.082 mmol) at 0° C. under nitrogen.The clear solution obtained was stirred at the same temperature for 5min and at room temperature for 2 hr. Additional pyrophosphoryl chloride(0.040 mL) was added and the mixture was stirred at room temperature for3 hr before water (0.3 mL) was added. The mixture was stirred at roomtemperature overnight. Purification using reverse phase HPLC (PhenomenexAXIA 5u 21.2×100 mm; gradient over 8 min from 20 to 100% of solvent B;solvent A: 0.1% TFA in water; solvent B: 0.1% TFA in acetonitrile),concentration, and lyophilization gave((1R,3S)-1-amino-3-((R)-2-((pentyloxy)methyl)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)cyclopentyl)methyldihydrogen phosphate (1.5 mg, 3.32 μmol) as a white solid. LC/MSM⁺¹=430. HPLC retention time=6.81 min (condition L) ¹H NMR (500 MHz,METHANOL-d₄) δ 6.84-6.69 (m, 3H), 4.32-4.21 (m, 2H), 4.06-3.97 (m, 1H),3.96-3.90 (m, 1H), 3.89-3.83 (m, 1H), 3.69-3.65 (m, 1H), 3.64-3.59 (m,1H), 3.51 (t, J=6.5 Hz, 2H), 3.15-3.06 (m, 1H), 2.47 (dd, J=13.0, 6.7Hz, 1H), 2.14-2.06 (m, 1H), 2.04-1.83 (m, 3H), 1.67 (t, J=12.8 Hz, 1H),1.62-1.55 (m, 2H), 1.37-1.32 (m, 4H), 0.95-0.88 (m, 3H).

The phosphate ester examples (R₁ is —OP(O)(OH)₂) in Table 28 wereprepared according to the general procedure for Example 412.

TABLE 28 HPLC Example Phosphate Ester Retention HPLC MS No. of ExampleNo. MW Time (min.) condition (M⁺¹) 413 2 429.5 6.75 L 430 414 5 415.42.55 C 416 415 6 415.4 2.55 C 416 416 8 424.5 6.09 L 425 417 10 424.56.08 L 425 418 14 411.5 7.09 L 412 419 15 411.5 3.53 C 412 420 16 411.53.53 C 412 421 19 411.5 3.37 C 412 422 20 411.5 3.50 C 412 423 21 411.53.56 C 412 424 22 411.5 3.56 C 412 425 23 411.5 3.66 C 412 426 24 411.53.38 C 412 427 25 411.5 3.38 C 412 428 26 425.5 9.97 L 426 429 27 425.59.97 L 426 430 28 411.5 7.08 L 412 431 29 411.5 7.09 L 412 432 31 439.50.94 G 440 433 32 411.0 0.82 G 412 434 33 411.0 0.82 G 412 435 34 425.58.82 L 426 436 37 397.4 0.77 G 398 437 38 423.4 3.51 B 424 438 39 423.43.32 B 424 439 41 423.5 7.19 L 424 440 42 437.5 3.79 B 438 441 45 431.56.63 L 432 442 46 461.8 6.59 L 462 443 47 461.8 6.59 L 462 444 48 475.57.07 L 476 445 50 445.5 8.18 L 446 446 52 411.5 7.02 L 412 447 53 411.57.03 L 412 448 54 425.5 8.81 L 426 449 57 449.5 7.34 L 450 450 63 495.612.57 L 496 451 64 465.5 8.20 L 466 452 65 409.5 6.51 L 410 453 66 423.57.04 L 424 454 67 423.5 7.20 L 424 455 68 423.5 7.11 L 424 456 69 423.57.07 L 424 457 70 425.5 3.40 C 426 458 71 427.5 5.44 L 428 459 72 431.48.28 L 432 460 73 473.5 7.77 L 474 461 74 437.5 7.61 L 438 462 75 437.57.69 L 438 463 76 439.4 6.38 L 440 464 77 439.5 3.53 C 440 465 78 441.55.84 L 442 466 79 445.4 7.59 L 446 467 80 445.4 8.93 L 446 468 81 445.48.85 L 446 469 82 445.4 8.85 L 446 470 83 445.4 8.78 L 446 471 84 449.56.00 L 450 472 85 451.5 6.78 L 452 473 86 451.5 6.68 L 452 474 88 459.57.47 L 460 475 89 461.5 2.97 C 462 476 90 461.5 3.13 C 462 477 91 462.43.93 L 463 478 94 473.5 8.52 L 474 479 95 473.5 8.61 L 474 480 96 475.50.80 G 476 481 97 475.5 7.02 L 476 482 99 475.5 6.95 L 476 483 100 475.57.05 L 476 484 102 479.4 8.24 L 480 485 103 479.4 7.12 L 480 486 104479.4 7.04 L 480 487 105 479.4 7.54 L 480 488 107 479.6 7.70 L 480 489108 479.6 7.79 L 480 490 109 488.5 4.29 L 489 491 110 491.5 2.89 C 492492 112 493.6 3.87 C 494 493 113 495.9 8.84 L 496 494 117 497.5 3.20 C498 495 118 502.5 5.75 L 503 496 119 503.5 8.20 L 504 497 120 515.5 8.27L 516 498 122 517.6 3.82 C 518 499 123 519.5 7.89 L 520 500 124 544.66.91 L 545 501 125 530.6 6.97 L 531 502 142 413.5 8.73 L 414 503 143413.5 8.76 L 414 504 144 427.5 3.42 C 428 505 145 427.5 0.88 G 428.1 506146 427.5 0.89 G 428.1 507 147 447.5 0.86 G 448.1 508 148 447.5 0.86 G448.1 509 149 427.5 3.42 C 428 510 150 441.6 3.61 C 442 511 151 441.63.60 C 442 512 152 441.6 3.64 C 442 513 153 441.6 3.65 C 442 514 154441.6 3.57 C 442 515 155 441.6 3.59 C 442 516 156 448.5 2.35 C 449 517157 448.5 1.64 C 449 518 158 455.6 3.69 C 456 519 159 455.6 3.72 C 456520 160 461.6 3.55 C 462 521 163 475.6 3.51 C 476 522 164 475.6 3.51 C476 523 165 475.6 3.79 C 476 524 166 476.6 1.84 C 477 525 167 476.6 1.88C 477 526 169 477.6 2.60 C 478 527 170 477.6 3.16 C 478 528 171 477.63.34 C 478 529 172 477.6 3.32 C 478 530 173 489.6 3.75 C 490 531 175507.6 3.10 C 508 532 177 413.5 3.22 C 414 533 178 413.5 3.24 C 414 534179 427.5 9.13 L 428 535 180 427.5 9.22 L 428 536 181 427.5 3.41 C 428537 182 427.5 3.44 C 428 538 183 441.6 3.56 C 442 539 176 441.6 3.55 C442 540 186 383.4 0.67 G 384 541 187 397.5 0.98 G 398 542 191 425.5 8.40L 426 543 192 425.5 8.36 L 426 544 193 475.5 7.47 L 476 545 194 475.57.48 L 476 546 195 459.5 8.58 L 460 547 196 459.5 3.61 B 460 548 197459.5 8.83 L 460 549 198 459.5 8.83 L 460 550 199 459.5 0.81 G 460 551200 459.5 7.71 L 460 552 201 459.5 0.84 G 460 553 202 475.5 0.77 G 476554 203 463.4 7.12 L 464 555 204 407.5 8.26 L 408 556 205 421.5 8.73 L422 557 206 451.5 7.12 L 452 558 207 437.5 6.57 L 438 559 208 473.5 8.05L 474 560 209 473.5 8.01 L 474 561 211 438.5 4.06 L 439 562 213 500.60.93 G 501 563 214 473.5 8.40 L 474 564 215 473.5 8.19 L 474 565 216445.5 0.85 G 446 566 217 445.5 0.83 G 446 567 221 445.5 0.83 G 446 568223 445.5 0.82 G 446 569 226 401.4 0.80 G 402 570 227 401.4 0.81 G 402571 228 415.5 7.32 L 416 572 229 415.5 7.31 L 416 573 230 429.5 0.89 G430 574 233 402.4 0.46 G 403 575 234 395.5 0.93 G 396 576 235 395.5 0.94G 396 577 236 381.5 0.88 G 396 578 237 381.5 0.88 G 396 579 238 425.52.46 B 426 580 240 453.5 7.54 L 454 581 243 475.5 7.75 L 476 582 244461.5 6.62 L 462 583 245 461.5 6.61 L 462 584 246 439.5 5.96 L 440 585247 467.5 6.80 L 468 586 250 455.5 6.95 L 456 587 251 455.5 6.86 L 456588 255 481.5 6.28 L 482 589 260 481.5 6.28 L 482 590 265 475.5 7.08 L476 591 266 441.5 7.50 L 442 592 267 441.5 7.50 L 442 593 268 459.5 0.88G 456 594 269 459.5 0.88 G 456 595 270 397.5 0.72 G 398 596 272 411.50.80 G 412 597 274 439.5 0.86 G 440 598 275 425.5 1.01 B 426 599 277425.5 1.01 B 426 600 278 425.5 8.39 L 426 601 281 423.5 0.80 G 424 602282 425.5 8.45 L 426 603 284 443.5 0.80 G 444 604 289 480.5 0.60 G 481605 290 480.5 0.60 G 481 606 292 430.5 0.64 G 431 607 299 477.5 0.93 G478 608 300 477.5 1.12 M 478 609 305 477.5 1.14 M 478 610 315 439.5 0.86G 440 611 316 439.5 1.10 M 440 612 318 410.5 0.74 G 411 613 327 444.55.64 L 445 614 328 444.5 5.61 L 445 615 332 472.5 6.00 L 473 616 334424.5 5.60 L 425 617 335 424.5 1.32 G 425 618 337 479.5 6.98 L 480 620343 535.4 10.58 L 536 621 345 444.0 0.81 G 445 622 346 423.5 10.11 L 424623 347 439.5 0.83 G 440 624 348 467.5 0.89 G 468 625 351 440.5 0.70 G441 626 352 468.5 0.79 G 469 627 353 445.5 0.75 G 446 628 354 454.5 0.74G 455 629 355 438.5 0.67 G 439 630 357 451.5 0.90 G 453 631 358 451.50.89 G 453 632 359 451.5 0.89 G 453 633 360 451.5 0.90 G 453 634 361395.4 0.67 G 396 635 364 395.4 0.75 G 396 636 367 407.5 8.46 L 408 637368 407.5 8.45 L 408 638 369 423.5 6.62 L 424 639 370 405.5 6.35 L 406640 371 423.5 6.63 L 424 641 372 457.5 7.56 L 458 642 373 435.5 6.34 L436 643 374 457.5 8.79 L 458 644 375 441.5 8.11 L 442 645 376 421.5 9.01L 422 646 377 409.5 8.16 L 410 647 378 405.5 8.16 L 406 648 379 407.58.53 L 408 649 380 407.5 8.53 L 408 650 381 407.5 3.71 C 408 651 383407.5 3.68 C 408 652 384 327.5 8.28 L 328 653 385 327.5 8.42 L 328 654386 327.5 8.52 L 328 655 387 327.5 9.51 L 328 656 388 407.5 3.70 C 408657 389 407.5 3.76 C 408 658 390 407.5 3.72 C 408 659 392 421.5 10.28 L422 660 393 421.5 10.13 L 422 661 395 421.5 10.60 L 422 662 399 458.50.83 G 459 663 400 458.5 0.83 G 459 664 402 472.5 7.37 L 473 665 403438.5 7.45 L 439 666 409 410.4 6.36 L 411

Example 667 ((1R,3S)-1-amino-3-((6S)-6-((phenylsulfinyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyl Dihydrogen Phosphate

To a stirred clear solution of((1R,3S)-1-amino-3-((S)-6-((phenylthio)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate (4 mg, 8.94 μmol), DMSO (0.013 mL, 0.179 mmol), andL-10-(−)-camphor sulfonic acid (10.38 mg, 0.045 mmol) in dichloromethane(0.5 mL) and methanol (0.2 mL) cooled with dry-ice was added 77% m-CPBA(2.003 mg, 8.94 μmol). The temperature was raised to 0° C. over 50 min.The mixture was stirred at 0° C. for 30 min and room temperature for 30min. The mixture was concentrated. Purification using reverse phase HPLC(Waters Xbridge C18 19×100 mm; gradient over 8 min from 20 to 100% ofsolvent B; solvent A: 10% MeOH: 90% H₂O: 0.1% TFA; solvent B: 90% MeOH,10% H₂O, 0.1% TFA), concentration, and lyophilization gave((1R,3S)-1-amino-3-((6S)-6-((phenylsulfinyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyl dihydrogenphosphate (3.6 mg, 7.38 μmol) as a white solid. LC/MS M⁺¹=464. HPLCretention time=6.43 min (Condition L)¹H NMR (400 MHz, METHANOL-d₄) δ7.73-7.67 (m, 2H), 7.63-7.56 (m, 3H), 7.08-6.96 (m, 3H), 3.99-3.87 (m,2H), 3.18-3.09 (m, 1H), 3.02 (ddd, J=13.0, 7.3, 5.4 Hz, 1H), 2.96-2.79(m, 3H), 2.73-2.56 (m, 1H), 2.49 (dd, J=13.4, 7.0 Hz, 1H), 2.41-2.11 (m,3H), 2.07-1.89 (m, 4H), 1.78-1.58 (m, 2H).

The example in Table 29 was prepared according to the general procedureof Example 667.

TABLE 29 Ex. HPLC HPLC MS No. Structure MW ret. time (min.) condition(M + 1) 668

443.5 2.20 C 444

Example 669((1R,3S)-1-amino-3-((S)-6-((phenylsulfonyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate

To a stirred clear solution of((1R,3S)-1-amino-3-((S)-6-((phenylthio)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate (4 mg, 8.94 μmol) and L-10-(−)-camphor sulfonicacid (10.38 mg, 0.045 mmol) in dichloromethane (0.5 mL) and methanol(0.2 mL) was added 77% m-CPBA (4.01 mg, 0.018 mmol). The mixture wasstirred at room temperature for 2 h before being concentrated.Purification using reverse phase HPLC (Waters Xbridge C18 19×100 mm;gradient over 8 min from 20 to 100% of solvent B; solvent A: 10% MeOH:90% H₂O: 0.1% TFA; solvent B: 90% MeOH, 10% H₂O, 0.1% TFA),concentration, and lyophilization gave((1R,3S)-1-amino-3-((S)-6-((phenylsulfonyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyldihydrogen phosphate (3.3 mg, 6.88 μmol) as a white solid. LC/MSM⁺¹=480. HPLC retention time=6.98 min (condition L ¹H NMR (400 MHz,METHANOL-d₄) δ 7.98-7.92 (m, 2H), 7.77-7.72 (m, 1H), 7.68-7.59 (m, 2H),6.99 (q, J=7.9 Hz, 3H), 3.98-3.84 (m, 2H), 3.25 (dd, J=6.4, 5.1 Hz, 2H),3.03-2.95 (m, 1H), 2.84-2.76 (m, 2H), 2.60 (dd, J=16.6, 9.6 Hz, 1H),2.48 (dd, J=13.1, 6.7 Hz, 1H), 2.37 (br. s., 1H), 2.18-1.87 (m, 6H),1.76-1.56 (m, 2H).

Examples 672 and 673((1R,3S)-1-amino-3-((S)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(672) and((1R,3S)-1-amino-3-((R)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(673)

Preparation 672A: 5-methoxypent-1-yne

To a mixture of pent-4-yn-1-ol (2 mL, 21.49 mmol) in THF (20 mL) wasadded sodium hydride (1032 mg, 25.8 mmol) portionwise over 15 minutes.The reaction mixture was stirred 30 minutes after addition and thenmethyl iodide (2.69 mL, 43.0 mmol) was added. The reaction mixture washeated at 40° C. for 6 h. An aliquot was removed, concentrated andchecked by NMR. Reaction was incomplete. Additional sodium hydride (1032mg, 25.8 mmol) and methyl iodide (2.69 mL, 43.0 mmol) were added and thereaction mixture was stirred overnight at 40° C. An aliquot check showreaction was complete. The reaction mixture was diluted with ethylacetate and washed with H₂O. The organic layer was dried with MgSO₄,filtered and concentrated. The crude product was distilled at 90 to 100°C. to give 5-methoxypent-1-yne (950 mg, 9.68 mmol) as a clear liquid.

Preparation 672B:(5R,7S)-7-(6-(5-methoxypent-1-yn-1-yl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-3,4-dihydronaphthalen-2-yltrifluoromethanesulfonate (1.35 g, 3.23 mmol), copper(I) iodide (0.062g, 0.323 mmol), and bis(triphenylphosphine)palladium(II) chloride (0.227g, 0.323 mmol) in TEA (3 mL) was added 5-methoxypent-1-yne (1.587 g,16.17 mmol). The reaction mixture was heated at 60° C. for 1 hour. Thereaction mixture was diluted with ethyl acetate and washed with 1M HCl.The organic layer was dried with MgSO₄, filtered and concentrated. Thecrude material was purified on a silica gel cartridge (40 g) using anEtOAc/Hex gradient (0-100% EtOAc over 12 CV). Fractions 28-33 wereisolated, concentrated, and dried in vacuo to afford(5R,7S)-7-(6-(5-methoxypent-1-yn-1-yl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(1 g, 2.74 mmol). HPLC retention time=1.00 min (condition A); LC/MSM⁺¹=346; ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.08-6.89 (m, 3H), 6.69 (s,1H), 5.62 (s, 1H), 4.44-4.22 (m, 2H), 3.52 (t, J=6.2 Hz, 2H), 3.39 (s,3H), 3.14-2.97 (m, 1H), 2.83 (t, J=8.1 Hz, 2H), 2.50 (t, J=7.2 Hz, 2H),2.46-2.39 (m, 2H), 2.33 (dd, J=13.2, 7.3 Hz, 1H), 2.21-2.09 (m, 2H),2.03-1.92 (m, 2H), 1.85 (quin, J=6.7 Hz, 3H).

Preparations 672C and 673C:(5R,7S)-7-((S)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(672C-Isomer 1) and(5R,7S)-7-((R)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(673-Isomer 2)

To a mixture of(5R,7S)-7-(6-(5-methoxypent-1-yn-1-yl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(105 mg, 0.287 mmol) in MeOH (10 mL) was added Pearlman's Catalyst(20.17 mg, 0.144 mmol). The reaction mixture was hydrogenated under aballoon of H₂ for 1 hour. LCMS show complete hydrogenation. The catalystwas removed by filtration. The mixture was concentrated in vacuo toafford 105 mg of desired product. The individual isomers were separatedusing a Chiral AD-H 25×3 cm ID, Sum under SFC conditions (30% MeOH inC02). Two fractions were obtained and concentrated to dryness. Peak 1:recovered(5R,7S)-7-((S)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(40 mg, 0.108 mmol). Peak 2: recovered(5R,7S)-7-((R)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(38 mg, 0.102 mmol).

Example 672

To a mixture of(5R,7S)-7-((S)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(40 mg, 0.108 mmol) in DMSO (0.5 mL) and MeOH (1 mL) was added 1N NaOH(0.5 mL). The reaction mixture was heated at 95° C. for 2 hours. Next,the mixture was cooled and then acidified with TFA. The mixture wasfiltered and purified by HPLC. HPLC conditions: Phenomenex Luna 5 micronC18 column (30×100 mm); MeCN (0.1% TFA)/water (0.1% TFA); 20%-100%gradient over 15 minutes; 30 mL/min. Fractions with correct mass wereisolated and freeze-dried overnight. Recovered((1R,3S)-1-amino-3-((S)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (32 mg, 0.067 mmol). HPLC retention time=8.15 min (condition L);LC/MS M⁺¹=346; ¹H NMR in CD₃OD (400 MHz, METHANOL-d₄) δ 7.05-6.94 (m,3H), 3.72-3.56 (m, 2H), 3.42 (t, J=6.5 Hz, 2H), 3.34 (s, 3H), 3.19-3.03(m, 1H), 2.91-2.70 (m, 3H), 2.50-2.28 (m, 2H), 2.20-2.04 (m, 1H),2.03-1.86 (m, 4H), 1.79-1.65 (m, 2H), 1.61 (quin, J=7.0 Hz, 2H),1.53-1.23 (m, 7H).

Example 673

To a mixture of(5R,7S)-7-((R)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(40 mg, 0.108 mmol) in DMSO (0.5 mL) and MeOH (1 mL) was added 1N NaOH(0.5 mL). The reaction mixture was heated at 95° C. for 2 hours. Themixture was cooled and then acidified with TFA. The mixture was filteredand purified by HPLC. HPLC conditions: Phenomenex Luna 5 micron C18column (30×100 mm); MeCN (0.1% TFA)/water (0.1% TFA); 20%-100% gradientover 15 minutes; 30 mL/min. Fractions with correct mass were isolatedand freeze-dried overnight. Recovered((1R,3S)-1-amino-3-((R)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (26 mg, 0.055 mmol). HPLC retention time=8.16 min (condition L);LC/MS M⁺¹=346; ¹H NMR in CD₃OD (400 MHz, METHANOL-d₄) δ 7.02-6.96 (m,3H), 3.71-3.56 (m, 2H), 3.42 (t, J=6.6 Hz, 2H), 3.34 (s, 3H), 3.18-3.02(m, 1H), 2.90-2.71 (m, 3H), 2.49-2.29 (m, 2H), 2.12 (d, J=2.9 Hz, 1H),2.02-1.87 (m, 4H), 1.79-1.66 (m, 2H), 1.66-1.55 (m, 2H), 1.52-1.32 (m,7H).

Examples 674 and 675((1R,3R)-1-amino-3-(6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 674A:(5R,7R)-7-(6-(5-methoxypent-1-yn-1-yl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-3,4-dihydronaphthalen-2-yltrifluoromethanesulfonate (130 mg, 0.311 mmol), copper(I) iodide (5.93mg, 0.031 mmol), and bis(triphenylphosphine)palladium(II) chloride(21.86 mg, 0.031 mmol) in TEA (311 μl) was added 5-methoxypent-1-yne(153 mg, 1.557 mmol). The mixture was stirred at 60° C. for 1 hour. LCMSshowed complete conversion. The reaction mixture was diluted with ethylacetate and washed with 1M HCl. The organic layer was dried with MgSO₄,filtered and concentrated under reduced pressure. The crude material waspurified on a silica gel cartridge (40 g) using an EtOAc/Hex gradient(0-100% EtOAc over 12 CV) giving access to(5R,7R)-7-(6-(5-methoxypent-1-yn-1-yl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(70 mg, 0.192 mmol). HPLC retention time=0.99 min (condition G) LC/MSM⁺¹=366.4. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.06-6.98 (m, 2H), 6.94 (d,J=7.7 Hz, 1H), 6.63 (s, 1H), 4.34 (dd, J=12.5, 7.9 Hz, 2H), 3.52 (t,J=6.3 Hz, 2H), 3.36 (s, 3H), 3.29-3.17 (m, 1H), 2.78 (t, J=8.1 Hz, 2H),2.46 (t, J=7.0 Hz, 2H), 2.40-2.31 (m, 2H), 2.26 (dd, J=13.3, 7.4 Hz,1H), 2.21-2.10 (m, 2H), 2.03-1.91 (m, 1H), 1.91-1.63 (m, 4H).

Preparation 674B:(5R,7R)-7-(6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of(5R,7R)-7-(6-(5-methoxypent-1-yn-1-yl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(70 mg, 0.192 mmol) in EtOH (1596 μl) and EtOAc (319 μl) was addedpalladium hydroxide on carbon (26.9 mg, 0.038 mmol) at room temperature.The reaction mixture was purged with H₂ and stirred under H₂ overnight.LCMS showed complete conversion. The suspension was filtered throughCelite and concentrated to provide(5R,7R)-7-(6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(70 mg, 0.188 mmol) as a white solid. HPLC retention time=1.12 min(condition G) LC/MS M⁺¹=418.3. The individual isomers were separatedusing a Chiral OJ-H 25×3 cm ID, Sum under SFC conditions (30% MeOH inCO₂). Two fractions were obtained and concentrated to dryness.

Examples 674 and 675:((1R,3R)-1-amino-3-(6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a solution of(5R,7R)-7-(6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(21 mg, 0.057 mmol) in dioxane (565 μl) was added NaOH (565 μl, 0.565mmol). The temperature was elevated to 98° C. and LCMS showed completeconversion after 2 h. The reaction mixture was diluted with EtOAc andthe aqueous layer was back-extracted with EtOAc. HPLC prep purification:HPLC: condition=2 mL injection, gradient time of 5 min, start B=20% to100%, stop time of 15 min, Solvent A=0.1% TFA in water, Solvent B=0.1%TFA in MeCN, column=LUNA, wavelength of 220 nm. The product was thenfree base by extraction DCM/1N NaOH affording((1R,3R)-1-amino-3-(6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(10 mg, 0.028 mmol).

Isomer 1. HPLC retention time=8.18 min (condition L); LC/MS M⁺¹=346.4.¹H NMR (400 MHz, METHANOL-d₄) δ 7.04-6.92 (m, 3H), 3.71-3.58 (m, 2H),3.42 (t, J=6.5 Hz, 2H), 3.39 (s, 3H), 3.31-3.25 (m, 1H), 2.88-2.72 (m,3H), 2.36 (dd, J=16.4, 10.5 Hz, 1H), 2.26-2.11 (m, 3H), 2.02-1.91 (m,1H), 1.89-1.73 (m, 3H), 1.73-1.66 (m, 1H), 1.66-1.53 (m, 2H), 1.52-1.32(m, 7H). Isomer 2: HPLC retention time=8.17 min (condition L); LC/MSM⁺¹=346.4; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.04-6.92 (m, 3H), 3.71-3.58(m, 2H), 3.42 (t, J=6.5 Hz, 2H), 3.39 (s, 3H), 3.31-3.25 (m, 1H),2.88-2.72 (m, 3H), 2.36 (dd, J=16.4, 10.5 Hz, 1H), 2.26-2.11 (m, 3H),2.02-1.91 (m, 1H), 1.89-1.73 (m, 3H), 1.73-1.66 (m, 1H), 1.66-1.53 (m,2H), 1.52-1.32 (m, 7H).

Examples 676 and 677((1R,3S)-1-amino-3-((S)-6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(676) and((1R,3S)-1-amino-3-((R)-6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(677)

Preparation 676A:(5R,7S)-7-(6-((3-methoxyphenyl)ethynyl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-3,4-dihydronaphthalen-2-yltrifluoromethanesulfonate (100 mg, 0.240 mmol), copper(I) iodide (4.56mg, 0.024 mmol), and bis(triphenylphosphine)palladium(II) chloride(16.82 mg, 0.024 mmol) in TEA (3 mL) was added1-ethynyl-3-methoxybenzene (0.091 mL, 0.719 mmol). The reaction mixturewas heated at 60° C. for 1 hour. The reaction mixture was diluted withethyl acetate and washed with 1M HCl. The organic layer was dried withMgSO₄, filtered and concentrated. The crude material was purified on asilica gel cartridge (12 g) using an EtOAc/Hex gradient (0-100% EtOAcover 20 CV). Fractions 15-17 were isolated, concentrated, and dried invacuo to afford(5R,7S)-7-(6-((3-methoxyphenyl)ethynyl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(68 mg, 0.170 mmol). HPLC retention time=1.09 min (condition A) LC/MSM⁺¹=400; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.30-7.22 (m, 1H), 7.14-6.97(m, 5H), 6.92 (ddd, J=8.4, 2.6, 0.9 Hz, 1H), 6.83 (s, 1H), 6.42-6.01 (m,1H), 4.84 (s, 3H), 4.41-4.25 (m, 2H), 3.33 (dt, J=3.2, 1.6 Hz, 2H), 3.06(tt, J=11.0, 7.2 Hz, 1H), 2.86 (t, J=8.1 Hz, 2H), 2.49 (td, J=8.1, 1.3Hz, 2H), 2.30 (dd, J=13.0, 7.3 Hz, 1H), 2.19-2.05 (m, 2H), 2.00-1.74 (m,3H).

Preparations 676B and 677B:(5R,7S)-7-((R)-6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(677B) and(5R,7S)-7-((S)-6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(677B)

To a mixture of(5R,7S)-7-(6-((3-methoxyphenyl)ethynyl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(68 mg, 0.170 mmol) in MeOH (3 mL) and ethyl acetate (1 mL) was addedPearlman's Catalyst (23.90 mg, 0.170 mmol). The reaction mixture washydrogenated under a balloon of H₂ for 1 hour. The mixture was filteredto remove the catalyst and concentrated in vacuo. The individual isomerswere separated using a Chiral AS-H 25×3 cm ID, 5 um under SFC conditions(37% MeOH in CO₂). Two fractions which were obtained and concentrated todryness. Isomer 1: recovered(5R,7S)-7-(6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(17 mg, 0.042 mmol). Isomer 2; recovered(5R,7S)-7-(6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(16 mg, 0.039 mmol).

Example 676

To a mixture of(5R,7S)-7-(6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(Isomer 1, 17 mg, 0.042 mmol) in MeOH (1 mL) and DMSO (0.5 mL) was added1N NaOH (1 mL). The reaction mixture was heated at 90° C. overnight, andthen cooled and acidified with TFA. The mixture was filtered andpurified by HPLC. HPLC conditions: Phenomenex Luna 5 micron C18 column(30×100 mm); MeCN (0.1% TFA)/water (0.1% TFA); 20%-100% gradient over 15minutes; 30 mL/min. Fractions with correct mass were isolated andfreeze-dried overnight. Recovered((1R,3S)-1-amino-3-(6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (14 mg, 0.028 mmol). HPLC retention time=8.91 min (condition L)LC/MS M⁺¹=380. ¹H NMR in CD₃OD (400 MHz, METHANOL-d₄) δ 7.18 (t, J=7.8Hz, 1H), 7.05-6.96 (m, 3H), 6.85-6.76 (m, 2H), 6.74 (dd, J=8.3, 1.9 Hz,1H), 3.79 (s, 3H), 3.71-3.56 (m, 2H), 3.19-3.02 (m, 1H), 2.89 (dd,J=16.4, 3.6 Hz, 1H), 2.83-2.75 (m, 2H), 2.75-2.68 (m, 2H), 2.50-2.35 (m,2H), 2.20-2.06 (m, 1H), 2.06-1.87 (m, 4H), 1.82-1.61 (m, 4H), 1.53-1.36(m, 1H).

Example 677

To a mixture of(5R,7S)-7-(6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(Isomer 2, 16 mg, 0.042 mmol) in MeOH (1 mL) and DMSO (0.5 mL) was added1N NaOH (1 mL). The reaction mixture was heated at 90° C. overnight, andthen cooled and acidified with TFA. The mixture was filtered andpurified by HPLC. HPLC conditions: Phenomenex Luna 5 micron C18 column(30×100 mm); MeCN (0.1% TFA)/water (0.1% TFA); 20%-100% gradient over 15minutes; 30 mL/min. Fractions with correct mass were isolated andfreeze-dried overnight. Recovered((1R,3S)-1-amino-3-(6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (14 mg, 0.028 mmol). HPLC retention time=8.89 min (condition L)LC/MS M⁺¹=380; ¹H NMR in CD₃OD (400 MHz, METHANOL-d₄) δ 7.18 (t, J=7.8Hz, 1H), 7.04-6.97 (m, 3H), 6.85-6.77 (m, 2H), 6.74 (dd, J=8.3, 1.9 Hz,1H), 3.79 (s, 3H), 3.71-3.57 (m, 2H), 3.20-3.02 (m, 1H), 2.89 (dd,J=16.5, 3.3 Hz, 1H), 2.83-2.75 (m, 2H), 2.75-2.69 (m, 2H), 2.50-2.36 (m,2H), 2.19-2.06 (m, 1H), 2.06-1.88 (m, 4H), 1.81-1.64 (m, 4H), 1.44 (dtd,J=12.8, 10.4, 5.9 Hz, 1H).

Examples 678 and 679((1R,3S)-1-amino-3-((R)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(678) and((1R,3S)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(679)

Preparation 678A:3-(6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propyl4-methylbenzenesulfonate

To a mixture of6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-3,4-dihydronaphthalen-2-yltrifluoromethanesulfonate (330 mg, 0.791 mmol), copper(I) iodide (15.06mg, 0.079 mmol), and bis(triphenylphosphine)palladium(II) chloride (55.5mg, 0.079 mmol) in TEA (3 mL) was added benzyl propargyl ether (0.572mL, 3.95 mmol). The reaction mixture was stirred at room temperature for1 hour. The reaction mixture was diluted with ethyl acetate and washedwith 1M HCl. The organic layer was dried with MgSO₄, filtered andconcentrated. The crude material was purified on a silica gel cartridge(40 g) using an EtOAc/Hex gradient (0-100% EtOAc over 12 CV). Fractions28-31 were isolated, concentrated, and dried in vacuo. The solidmaterial was dissolved in MeOH (10 mL) and Pearlman's Catalyst (111 mg,0.791 mmol) was added. The reaction mixture was hydrogenated underballoon pressure for 18 hours. The mixture was filtered to remove thecatalyst and concentrated in vacuo. The solids were dissolved pyridine(5 mL) and then p-toluenesulfanonyl chloride (452 mg, 2.372 mmol) wasadded. After 2 hours, additional p-toluenesulfanonyl chloride (452 mg,2.372 mmol) was added. The reaction mixture was diluted with ethylacetate and washed with H₂O. The organic layer was dried with MgSO₄,filtered and concentrated. The crude material was purified on a silicagel cartridge (40 g) using an EtOAc/Hex gradient (0-100% EtOAc over 20CV). Fractions 20-22 were concentrated and dried in vacuo to afford3-(6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propyl4-methylbenzenesulfonate (200 mg, 0.414 mmol). ¹H NMR (400 MHz,CHLOROFORM-d) δ 7.82 (d, J=8.1 Hz, 2H), 7.37 (d, J=7.9 Hz, 2H),7.05-6.89 (m, 3H), 5.54 (s, 1H), 4.41-4.24 (m, 2H), 4.08 (t, J=6.5 Hz,2H), 3.11-2.95 (m, 1H), 2.77 (td, J=10.0, 5.4 Hz, 3H), 2.47 (s, 3H),2.40-2.26 (m, 2H), 2.21-2.08 (m, 2H), 2.03-1.73 (m, 6H), 1.70-1.55 (m,2H), 1.48-1.32 (m, 2H).

Preparations 678B and 679B:(5R,7S)-7-((R)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(678B) and(5R,7S)-7-((R)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(679B)

To ethanol (0.5 mL, 0.248 mmol) was added sodium (114 mg, 4.96 mmol).The mixture was stirred until the sodium metal was consumed. A solutionof3-(6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propyl4-methylbenzenesulfonate (120 mg, 0.248 mmol) in DMF was added and thereaction mixture was stirred at room temperature. The reaction mixturewas stirred for 12 hours. The reaction mixture was diluted with ethylacetate and washed with saturated NaCl. The organic layer was dried withMgSO₄, filtered and concentrated. The crude material was purified on asilica gel cartridge (12 g) using an EtOAc/Hex gradient (0-100% EtOAcover 21 CV. Recovered 65 mg of a mixture of isomers. The individualisomers were separated using a Chiral AS-H 25×3 cm ID, 5 um under SFCconditions (27% MeOH in CO₂). Two fractions which were obtained andconcentrated to dryness. Isomer 1: Recovered(5R,7S)-7-((R)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(30 mg, 0.084 mmol). Isomer 2: Recovered(5R,7S)-7-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(31 mg, 0.087 mmol).

Example 678

To a mixture of(5R,7S)-7-((R)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(30 mg, 0.084 mmol) in DMSO (1 mL) and MeOH (1 mL) was added 1N NaOH.The reaction mixture was heated at 95° C. overnight. The mixture wascooled and acidified with TFA. The mixture was filtered and purified byHPLC. HPLC conditions: Phenomenex Luna 5 micron C18 column (30×100 mm);MeCN (0.1% TFA)/water (0.1% TFA); 20%-100% gradient over 15 minutes; 30mL/min. Fractions with the correct mass were isolated and freeze-driedovernight. The material was poured into 1N NaOH (50 mL), stirred for 1hour, and extracted with EtOAc. The organic layer was dried with MgSO₄,filtered, and concentrated to afford((1R,3S)-1-amino-3-((R)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(21 mg, 0.060 mmol). HPLC retention time=6.57 min (condition L); LC/MSM⁺¹=332; ¹H NMR in CD₃OD (400 MHz, METHANOL-d₄) δ 7.10-6.78 (m, 3H),3.61-3.40 (m, 6H), 3.02 (tt, J=11.2, 7.0 Hz, 1H), 2.90-2.70 (m, 3H),2.37 (dd, J=16.3, 10.3 Hz, 1H), 2.23 (dd, J=13.1, 7.6 Hz, 1H), 2.09-1.85(m, 3H), 1.85-1.64 (m, 5H), 1.61-1.51 (m, 1H), 1.49-1.31 (m, 3H), 1.21(t, J=7.0 Hz, 3H).

Example 679

To a mixture of(5R,7S)-7-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(30 mg, 0.084 mmol) in DMSO (1 mL) and MeOH (1 mL) was added 1N NaOH.The reaction mixture was heated at 95° C. overnight. The mixture wascooled, acidified with TFA, filtered, and purified by HPLC. HPLCconditions: Phenomenex Luna 5 micron C18 column (30×100 mm); MeCN (0.1%TFA)/water (0.1% TFA); 20%-100% gradient over 15 minutes; 30 mL/min.Fractions with correct mass were isolated and freeze-dried overnight.The material was poured into 1N NaOH (50 mL), stirred for 1 hour, andextracted with EtOAc (x). The organic layer was dried with MgSO₄,filtered, and concentrated to afford((1R,3S)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol (18 mg, 0.049 mmol). HPLC retention time=6.57 min (conditionL); LC/MS M⁺¹=332; ¹H NMR in CD₃OD (400 MHz, METHANOL-d₄) δ 7.20-6.76(m, 3H), 3.59-3.43 (m, 6H), 3.03 (tt, J=11.2, 7.0 Hz, 1H), 2.90-2.67 (m,3H), 2.36 (dd, J=16.2, 10.5 Hz, 1H), 2.26 (dd, J=12.8, 7.0 Hz, 1H),2.09-1.88 (m, 3H), 1.88-1.74 (m, 2H), 1.74-1.64 (m, 3H), 1.59 (t, J=12.4Hz, 1H), 1.50-1.28 (m, 3H), 1.20 (t, J=7.0 Hz, 3H).

Example 680((1R,3R)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 680A:(5R,7R)-7-((R)-6-(but-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of((R)-6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (250 mg, 0.549 mmol) and copper(I) bromide (157mg, 1.098 mmol) in THF (5 mL) was added allylmagnesium bromide (1100 μl,10.98 mmol) at room temperature and stirred at room temperature over 16h. The reaction mixture was diluted with saturated NH₃Cl and water andextracted with EtOAc. The organic layer was collected, dried overNa₂SO₄, concentrated on the rotavapor to give(5R,7S)-7-((S)-6-(4-(dimethylamino)benzyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one,LC/MS M⁺¹=326.

Preparation 680B:3-((S)-6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propanal

To a solution of(5R,7R)-7-((R)-6-(but-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(220 mg, 0.676 mmol) in THF (15 mL) was added NMO (158 mg, 1.352 mmol)and osmium tetroxide (6.37 μl, 0.020 mmol) at room temperature andstirred at room temperature over 16 h. Sodium periodate (578 mg, 2.70mmol) in H₂O (1 mL) was added and precipitate formed. The mixture wasstirred vigorously at room temperature under nitrogen for 30 min. Thereaction mixture was diluted with saturated NH₄Cl and water andextracted with EtOAc. The organic layer was collected, dried overNa₂SO₄, concentrated on the rotavapor to give3-((S)-6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propanal,LC/MS M⁺¹=328.

Preparation 680C:(5R,7R)-7-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of3-((S)-6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propanal(200 mg, 0.611 mmol), ethoxytrimethylsilane (361 mg, 3.05 mmol),triethylsilane (355 mg, 3.05 mmol) in nitromethane (2 mL) was addediron(III) chloride (9.91 mg, 0.061 mmol) at 0° C., and stirred at roomtemperature for 16. The mixture was filtered and purified by prep HPLCto give(5R,7R)-7-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one,LC/MS M⁺¹=358.

Example 680

To a solution of the crude(5R,7R)-7-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-onewhich was derived from previous step in dioxane (3 mL) and water (1 mL)was added LiOH (15.77 mg, 0.659 mmol), and stirred at 100° C. for 16 h.The reaction mixture was diluted with water, extracted with EtOAc. Theorganic layer was collected, dried over Na₂SO₄, and concentrated on therotavapor to give the crude product which was purified with preparativeHPLC: column Phenomenex Luna C18 5u 21.2×100 mm. Solvent A: 10% MeOH-90%H₂O-0.1% TFA; Solvent B: 90% MeOH-10% H₂O-0.1% TFA. Gradient time=15min. Start B=0%, Final B 100%. Stop time 25 min.((1R,3S)-1-amino-3-((S)-6-((Z)-hex-2-en-1-yloxy)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,LC/MS M⁺¹=332. HPLC method: L; HPLC ret. time 6.86 (min.). ¹H NMR (400MHz, METHANOL-d₄) δ 7.00-6.90 (m, 3H), 3.60-3.50 (m, 6H), 2.80-2.60 (m,3H), 2.41-1.80 (m, 6H), 1.78-1.30 (m, 9H), 1.24 (t, J=7.0 Hz, 3H).

Examples 681 and 682((1R,3S)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(681) and((1R,3S)-1-amino-3-((R)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(682)

Preparation 681A:(5R,7S)-7-(6-((2-methoxyphenyl)ethynyl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-3,4-dihydronaphthalen-2-yltrifluoromethanesulfonate (64 mg, 0.153 mmol), copper(I) iodide (2.92mg, 0.015 mmol), and bis(triphenylphosphine)palladium(II) chloride(10.76 mg, 0.015 mmol) in TEA (3 mL) was added1-ethynyl-2-methoxybenzene (0.059 mL, 0.460 mmol). The reaction mixturewas heated at 60° C. for 1 hour. The reaction mixture was diluted withethyl acetate and washed with 1M HCl. The organic layer was dried withMgSO₄, filtered and concentrated. The crude material was purified on asilica gel cartridge (24 g) using an EtOAc/Hex gradient (0-100% EtOAcover 12 CV). Fractions 20-23 were isolated, concentrated, and dried invacuo to afford(5R,7S)-7-(6-((2-methoxyphenyl)ethynyl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(45 mg, 0.113 mmol). HPLC retention time=1.07 min (condition A) LC/MSM⁺¹=400

Preparations 681B and 682B:(5R,7S)-7-(6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of(5R,7S)-7-(6-((2-methoxyphenyl)ethynyl)-7,8-dihydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(45 mg, 0.113 mmol) in MeOH (5 mL) was added Pearlman's Catalyst (15.82mg, 0.113 mmol). The reaction mixture was hydrogenated under a balloonof H₂ overnight. The mixture was filtered to remove the catalyst andthen concentrated in vacuo to afford 45 mg of a mixture of isomers. Theindividual isomers were separated using a Chiral OJ-H 25×3 cm ID, Sumunder SFC conditions (35% MeOH in CO₂). Two fractions which wereobtained and concentrated to dryness. Isomer 1: Recovered(5R,7S)-7-(6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(14 mg, 0.035 mmol). Isomer 2: Recovered(5R,7S)-7-(6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(14 mg, 0.035 mmol).

Example 681

To a mixture of(5R,7S)-7-(6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(40 mg, 0.108 mmol) in DMSO (0.5 mL) and MeOH (1 mL) was added 1N NaOH(0.5 mL). The reaction mixture was heated at 95° C. for 4 hours, cooled,and then acidified with TFA. The mixture was filtered and purified byHPLC. HPLC conditions: Phenomenex Luna 5 micron C18 column (30×100 mm);MeCN (0.1% TFA)/water (0.1% TFA); 20%-100% gradient over 15 minutes; 30mL/min. Fractions with the correct mass were isolated and freeze-driedovernight to afford((1R,3S)-1-amino-3-((R)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol, TFA (26 mg, 0.055 mmol). HPLC retention time=8.87min (condition L); LC/MS M⁺¹=380; ¹H NMR in CD₃OD (400 MHz, METHANOL-d₄)δ 7.21-7.10 (m, 2H), 7.05-6.96 (m, 3H), 6.91 (d, J=7.9 Hz, 1H), 6.86(td, J=7.4, 1.0 Hz, 1H), 3.83 (s, 3H), 3.72-3.54 (m, 2H), 3.20-3.03 (m,1H), 2.97-2.69 (m, 5H), 2.48-2.35 (m, 2H), 2.21-1.86 (m, 5H), 1.80-1.57(m, 4H), 1.52-1.36 (m, 1H).

Example 682

To a mixture of(5R,7S)-7-(6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(40 mg, 0.108 mmol) in DMSO (0.5 mL) and MeOH (1 mL) was added 1N NaOH(0.5 mL). The reaction mixture was heated at 95° C. for 4 hours, cooled,and then acidified with TFA. The mixture was filtered and purified byHPLC. HPLC conditions: Phenomenex Luna 5 micron C18 column (30×100 mm);MeCN (0.1% TFA)/water (0.1% TFA); 20%-100% gradient over 15 minutes; 30mL/min. Fractions with the correct mass were isolated and freeze-driedovernight to afford((1R,3S)-1-amino-3-((R)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (26 mg, 0.055 mmol). HPLC retention time=8.97 min (condition L);LC/MS M⁺¹=380; MS (m+1)=380; ¹H NMR in CD₃OD (400 MHz, METHANOL-d₄) δ7.21-7.10 (m, 2H), 7.04-6.97 (m, 3H), 6.91 (d, J=7.9 Hz, 1H), 6.86 (td,J=7.4, 1.1 Hz, 1H), 3.83 (s, 3H), 3.72-3.54 (m, 2H), 3.19-3.03 (m, 1H),2.96-2.70 (m, 5H), 2.49-2.35 (m, 2H), 2.20-1.87 (m, 5H), 1.82-1.57 (m,4H), 1.43 (dtd, J=12.8, 10.5, 6.1 Hz, 1H).

Example 683((1R,3R)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 683A:(5R,7R)-7-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

The Grignard reagent (2-methoxybenzyl)magnesium chloride (2195 μl, 0.549mmol) was added to a stirred mixture of((R)-6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (50 mg, 0.110 mmol) and copper(I) bromide (31.5mg, 0.220 mmol) in THF (10 ml) at −78° C. under nitrogen. The mixturewas stirred at −78° C. and was slowly raised to room temperature andstirred for 2 days. The mixture was heated at 60° C. for another 6 h.The reaction mixture was cooled down to 0° C. Next, 1 ml of water wasadded and the mixture was mixed with EtOAc (30 ml) and water (20 ml).The organic phase was separated and washed with saturated NH₄Cl (2×20ml) and brine (20 ml). The organic solution was dried over anhydroussodium sulfate and concentrated. Flash chromatography purification usingISCO (24 g silica gel column, gradient elution from 0 to 60% of EtOAc inhexane) to provide(5R,7R)-7-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.LC/MS M⁺¹=406.

Preparation 683B:((1R,3R)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

(5R,7R)-7-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-onewas mixed with 1,4-dioxane (2 ml), water (0.5 ml) and lithium hydroxidehydrate (69.1 mg, 1.646 mmol). The mixture was stirred at 100° C.overnight under N₂. The mixture was cooled and filtered, the filtratewas concentrated under vacuo and the residue was dissolved in DCM (20ml), washed with water (5 ml), dried (Na₂SO₄) and concentrated undervacuo. The residue was freeze dried to afford((1R,3R)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(25 mg over two steps). LC/MS M⁺¹=380. HPLC condition: L; HPLC ret. time7.84 (min.). ¹H NMR (400 MHz, METHANOL-d₄) δ 7.24-7.09 (m, 2H),7.03-6.80 (m, 5H), 3.83 (s, 3H), 3.59-3.42 (m, 2H), 2.98-2.67 (m, 5H),2.40 (dd, J=15.4, 10.6 Hz, 1H), 2.26-1.83 (m, 4H), 1.79-1.54 (m, 6H),1.50-1.25 (m, 2H).

Example 684((1R,3S)-1-amino-3-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 684A:(5R,7S)-7-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To the mixture of 3-methoxyphenol (123 mg, 0.988 mmol) in dry DMF (3ml), potassium tert-butoxide (790 μl, 0.790 mmol) in t-BuOH (lM) wasadded. After stirring at room temperature for 30 min,((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (90 mg, 0.198 mmol) in DMF (2 ml) was added andthe mixture was stirred at 65° C. for 4 h. The reaction was quenchedwith water (5 ml) at 0° C. The mixture was taken up in EtOAc (30 ml),washed with saturated NaHCO₃(3×20 ml), dried (Na₂SO₄) and concentratedunder vacuo. The residue was subject to flash chromatographypurification (12 g silica gel column, gradient elution from 0 to 70%ethyl acetate in hexanes, gradient time=18 min, out at 45% EtOAc) toafford (5R,7S)-7-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(70 mg). LC/MS M⁺¹=408.

Example 684

(5R,7S)-7-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(25 mg, 0.061 mmol) in dioxane (2 ml) was mixed with water (0.5 ml) andwas added lithium hydroxide hydrate (25.7 mg, 0.613 mmol), the mixturewas stirred at 100° C. for 16 h under N₂. After cooling, the mixture wasfiltered and washed with MeOH, the combined solvents were evaporated andthe residue was purified with preparative HPLC: column Phenomenex LunaC18 5u 21.2×100 mm. Solvent A: 10% MeOH-90% H₂O-0.1% TFA; Solvent B: 90%MeOH-10% H₂O-0.1% TFA. Gradient time=15 min. Start B=0%, Final B 100%.Stop time 20 min. The collected fraction was basified with saturatedNaHCO₃, concentrated under vacuo and the aqueous layer was extractedwith DCM (3×20 ml) which was dried (Na₂SO₄) and concentrated under vacuoto give ((1R,3S)-1-amino-3-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol (15 mg)as white solid. LC/MS M⁺¹=382. HPLC retention time=7.19 minutes(Condition L) ¹H NMR (400 MHz, METHANOL-d₄) δ 7.25-7.12 (m, 1H),7.07-6.96 (m, 3H), 6.59-6.47 (m, 3H), 3.94 (d, J=6.4 Hz, 2H), 3.79 (s,3H), 3.65-3.48 (m, 2H), 3.15-2.80 (m, 4H), 2.59 (dd, J=16.4, 10.5 Hz,1H), 2.40-1.79 (m, 7H), 1.70-1.51 (m, 2H).

Example 685((1R,3S)-1-amino-3-((S)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Example 685 was prepared according to the general procedure for Example684 using(5R,7S)-7-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one.MW 381.2; MS (M⁺¹)=382; HPLC method L, HPLC ret. time: 8.20 min.

Examples 686 and 687((1R,3R)-1-amino-3-(6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

Preparation 686A:(E)-2-(4-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)benzylidene)succinic Acid

To a mixture of K₂CO₃ (0.980 g, 7.09 mmol),(5R,7R)-7-(4-bromophenyl)-3-oxa-1-azaspiro[4.4]nonan-2-one (1.5 g, 5.06mmol), and itaconic acid (0.857 g, 6.58 mmol) in acetonitrile (14.98 ml)was slowly added water (4.50 ml). The mixture was stirred until theevolution of CO₂ stopped and then bubbled with nitrogen for 5 min.Palladium(II) acetate (0.057 g, 0.253 mmol) and tri-o-tolylphosphine(0.154 g, 0.506 mmol) were then added. Nitrogen was bubbled through thereaction mixture for 10 more minutes. The reaction mixture was heated at85° C. overnight with a reflux condenser. The reaction was completeaccording to LCMS. The reaction mixture was diluted with ethyl acetateand washed with 1N NaOH twice. The aqueous layers were combined andacidified with concentrated HCl to pH was 1-2. The aqueous layer wasextracted with EtOAC several times. The organic layers were combined,dried with Na₂SO₄, filtered, and concentrated under reduced pressure toafford(E)-2-(4-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)benzylidene)succinicacid (1.907 g, 5.52 mmol). HPLC retention time=0.63 min (condition G)LC/MS M⁺¹=346.3.

Preparation 686B:2-(4-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)benzyl)succinic Acid

To a mixture of (E)-2-(4-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)benzylidene)succinic acid (1.75 g, 5.07 mmol) in MeOH (100 mL) was addedPearlman's Catalyst (0.356 g, 0.507 mmol). The reaction mixture wasstirred under an atmosphere of H₂ overnight. LCMS showed completeconversion. The catalyst was removed by filtration through celite and2-(4-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)benzyl)succinic acid(2 g, 5.76 mmol) was obtained after concentration under reducedpressure. HPLC retention time=0.63 min (condition G) LC/MS M⁺¹=348.3.

Preparation 686C: Methyl4-oxo-6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate

To sulfuric acid (30 mL) was added2-(4-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)benzyl)succinic acid(1.761 g, 5.07 mmol). The solution was stirred overnight at roomtemperature. LCMS showed complete conversion. The solution was cooled to0° C. followed by the dropwise addition of MeOH (25.4 ml). After 2hours, the reaction was complete as judged by LCMS. The reaction mixturewas poured onto ice and the aqueous layer was extracted with EtOACseveral times until aqueous layer showed no desired product as judged byLCMS. The resulting solid was purified by ISCO using 100% hexanes to100% EtOAc as eluent affording methyl4-oxo-6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(1.54 g, 4.48 mmol). HPLC retention time=0.74 min (condition G) LC/MSM⁺¹=344.3. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.89 (d, J=1.8 Hz, 1H), 7.38(dd, J=7.9, 2.0 Hz, 1H), 7.26 (d, J=7.7 Hz, 1H), 6.48 (br. s., 1H),4.45-4.32 (m, 3H), 3.79-3.71 (m, 3H), 3.37-3.16 (m, 4H), 3.05-2.91 (m,1H), 2.91-2.81 (m, 1H), 2.43 (dd, J=13.6, 7.5 Hz, 1H), 2.35-2.22 (m,1H), 2.22-2.14 (m, 1H), 2.11-2.00 (m, 1H), 1.94-1.82 (m, 1H), 1.82-1.70(m, 1H).

Preparation 686D: Methyl6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate

To a solution of methyl4-oxo-6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(1.54 g, 4.48 mmol) in EtOH (44.8 ml) was added Pd(OH)₂ (0.630 g, 0.448mmol). The reaction mixture was placed under a hydrogen atmosphereovernight. LCMS showed complete conversion. The mixture was filteredthrough celite to remove the catalyst and the solution was concentratedunder reduced pressure to afford methyl6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(1.45 g, 4.40 mmol). HPLC retention time=0.87 min (condition G) LC/MSM⁺¹=330.3.

Preparation 686E:(5R,7R)-7-(6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a mixture of methyl6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(1.35 g, 4.10 mmol) in THF (100 mL) was added lithium borohydride in THF(4.10 ml, 8.20 mmol). The reaction mixture was heated at 60° C.overnight. The mixture was cooled and the reaction was quenched withwater. The reaction mixture was diluted with ethyl acetate and washedwith H₂O. The organic layer was dried with MgSO₄, filtered, andconcentrated to afford(5R,7R)-7-(6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(1 g, 3.32 mmol). HPLC retention time=0.74 min (condition G) LC/MSM⁺¹=302.1; Separation of Isomers:

Instrument: Berger SFC MGIII; SFC Prep Conditions Column: ChiralPak AD-H3×25 cm, Sum; Column Temp. 40° C.; Flow rate: 200 ml/min; Mobile Phase:CO₂/MEOH=60/40; Injection Program: Stacked (2.5 min/Cycle); SamplerConc. (mg/mL): 40 mg/mL; Detector Wavelength: 220 nm.

Isomer 1: HPLC retention time=0.74 min (condition G) LC/MS M⁺¹=302.1. ¹HNMR (400 MHz, CHLOROFORM-d) δ 7.07 (d, J=7.7 Hz, 1H), 6.99-6.90 (m, 2H),5.50 (br. s., 1H), 4.35 (q, J=8.5 Hz, 2H), 3.66 (dd, J=6.3, 1.7 Hz, 2H),3.25-3.09 (m, 1H), 2.95-2.77 (m, 3H), 2.50 (dd, J=16.3, 10.8 Hz, 1H),2.39 (dd, J=13.6, 7.5 Hz, 1H), 2.27-2.12 (m, 2H), 2.10-1.93 (m, 3H),1.87 (dd, J=13.6, 11.0 Hz, 1H), 1.81-1.70 (m, 1H), 1.53-1.38 (m, 2H).

Isomer 2: HPLC retention time=0.74 min (condition G) LC/MS M⁺¹=302.1.

Preparations 686F and 687F:(6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate

(5R,7R)-7-(6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(Isomer 1 of Preparation 686E, 410 mg, 1.360 mmol) was dissolved in drypyridine (1360 μl) and p-toluenesulfonyl chloride (519 mg, 2.72 mmol)was added in one portion. The resulting mixture was reacted at roomtemperature for 3 h. The solvent was removed in vacuo. The residue wasdissolved in DCM and loaded onto column with plenty DCM (to preventproduct crystallization on column). Flash chromatography purificationusing ISCO (40 g silica gel column, 20->100% ethyl acetate in hexanes)afforded(6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (550 mg, 1.207 mmol).

Preparation 686F (Isomer 1): HPLC retention time=1.00 min (condition G)LC/MS M⁺¹=456.1.

Preparation 687F (Isomer 2): HPLC retention time=0.99 min (condition G)LC/MS M⁺¹=456.1. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.88-7.78 (m, J=8.4Hz, 2H), 7.43-7.33 (m, J=7.9 Hz, 2H), 7.07-6.98 (m, 1H), 6.94 (d, J=7.9Hz, 1H), 6.90 (s, 1H), 5.20 (br. s., 1H), 4.41-4.25 (m, 2H), 4.02 (dd,J=6.6, 2.0 Hz, 2H), 3.23-3.07 (m, 1H), 2.92-2.73 (m, 3H), 2.48 (s, 3H),2.48-2.28 (m, 2H), 2.28-2.10 (m, 3H), 2.06-1.92 (m, 2H), 1.85 (dd,J=13.6, 11.2 Hz, 1H), 1.78-1.65 (m, 1H), 1.60-1.55 (m, 1H), 1.50-1.36(m, 1H).

Examples 686 and 687

To a suspension of(6-((5R,7R)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (Preparation 686F, 0.030 g, 0.066 mmol) indioxane (0.5 mL) was added 3-methoxyphenol (0.108 ml, 0.988 mmol)followed by potassium tert-butoxide (0.074 g, 0.659 mmol) at roomtemperature. The mixture was then heated at 70° C. for 2 h when LCMSshowed complete consumption of starting material. To this solution wasadded NaOH (0.5 mL, 0.500 mmol) at room temperature. The mixture washeated to 100° C. overnight. LCMS showed complete consumption ofstarting material. The solution was injected on the HPLC prep:condition=2 mL injection, gradient time of 5 min, start B=20% to 100%,stop time of 15 min, Solvent A=0.1% TFA in water, Solvent B=0.1% TFA inMeCN, column=LUNA, wavelength of 220 nm.((1R,3R)-1-amino-3-(6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,TFA (21 mg, 0.040 mmol) was obtained as a white solid with >95% purity.

Example 687 was prepared from Preparation 687F according to the generalprocedure of Example 686.

Example 686 (Isomer 1): HPLC retention time=8.19 min (condition L) LC/MSM⁺¹=382.1; H NMR (400 MHz, METHANOL-d₄) δ 7.22-7.12 (m, 1H), 7.09-7.03(m, 1H), 7.03-6.97 (m, 2H), 6.59-6.48 (m, 3H), 3.94 (d, J=6.4 Hz, 2H),3.79 (s, 3H), 3.65 (dd, J=11.9, 6.4 Hz, 2H), 2.96 (dd, J=16.6, 5.0 Hz,1H), 2.90-2.82 (m, 2H), 2.60 (dd, J=16.3, 10.3 Hz, 1H), 2.33-2.14 (m,4H), 2.11 (s, 1H), 1.90-1.72 (m, 3H), 1.61 (s, 1H).

Example 687 (Isomer 2): HPLC retention time=8.17 min (condition L) LC/MSM⁺¹=382.1; ¹H NMR (400 MHz, METHANOL-d₄) δ 7.22-7.12 (m, 1H), 7.09-7.03(m, 1H), 7.03-6.97 (m, 2H), 6.59-6.48 (m, 3H), 3.94 (d, J=6.4 Hz, 2H),3.79 (s, 3H), 3.65 (dd, J=11.9, 6.4 Hz, 2H), 2.96 (dd, J=16.6, 5.0 Hz,1H), 2.90-2.82 (m, 2H), 2.60 (dd, J=16.3, 10.3 Hz, 1H), 2.33-2.14 (m,4H), 2.11 (s, 1H), 1.90-1.72 (m, 3H), 1.61 (s, 1H).

PHOSPHORYLATED EXAMPLES Example 688((1R,3S)-1-amino-3-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methylDihydrogen Phosphate

To a mixture of((1R,3S)-1-amino-3-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(1.5 mg, 3.93 μmol) in MeCN (1 ml) was added pyridine (15.90 μl, 0.197mmol) and pyrophosphoryl chloride (14.85 mg, 0.059 mmol) at roomtemperature. The mixture was stirred at room temperature for 2 h. Water(0.5 ml) was added at 0° C. and the mixture was stirred at roomtemperature for 1 5 min. The mixture was purified with preparative(HPLC: column Phenomenex Luna C18 5u 21.2×100 mm. Solvent A: 10%MeOH-90% H₂O-0.1% TFA; Solvent B: 90% MeOH-10% H₂O-0.1% TFA. Gradienttime=15 mi. Start B=0, Final B 100. Stop time 25 min.) to afford 1 mg of((1R,3S)-1-amino-3-((R)-6-((3-methoxyphenoxy)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methyl dihydrogen phosphate, LC/MS M⁺¹=462. PLC Rt=7.07 min. (ConditionL). ¹H NMR (400 MHz, METHANOL-d₄) δ 7.17 (t, J=8.4 Hz, 1H), 7.04 (d,J=2.9 Hz, 3H), 6.60-6.46 (m, 3H), 4.04-3.86 (m, 4H), 3.79 (s, 3H), 3.15(s, 1H), 3.00-2.81 (m, 2H), 2.66-2.48 (m, 2H), 2.33-1.90 (m, 8H),1.82-1.68 (n, 1H).

The following compounds were prepared according to the generalprocedures of Example 688

HPLC Ex. ret. time HPLC MS No. Structure MW (min.) condition (M⁺¹)Comment 689   690

425.5   425.5 0.81   0.82 G   G 426   426 (S) Isomer 1 (R) Isomer 2 691692

425.5 425.5 1.01 1.01 B B 426 426 Isomer 1 Isomer 2 693   694

459.5   459.5 0.87   0.86 G   G 460   460 (S) Isomer 1 (R) Isomer 2 695  696

411.5   411.5 0.77   0.77 G   G 412   412 (R) Isomer 1 (S) Isomer 2 697  698

459.5   459.5 0.88   0.88 G   G 460   460 (S) Isomer 1 (R) Isomer 2 699

459.5 7.79 L 460 (S) Isomer 700

461.5 8.21 L 462 (S) Isomer 701 702

461.5 461.5 0.82 0.82 G G 462 462 Isomer 1 Isomer 2

Alternative Preparation of Example 672

To a stirred mixture of magnesium (1.814 g, 74.6 mmol) and anhydroustetrahydrofuran (3 mL) was added several drops of 1,2-dibromoethane atroom temperature under nitrogen. The mixture was stirred for 15 minbefore a solution of 1-bromo-4-methoxybutane (9.76 mL, 74.6 mmol) inanhydrous tetrahydrofuran (47 mL) was added dropwise to keep thereaction mixture warm but not boiling. After the addition, the mixturewas stirred at 60° C. under nitrogen for 3 hr. The solution wasseparated and added to a stirred mixture of copper(I) bromide (1.071 g,7.46 mmol),((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (1.7 g, 3.73 mmol) and tetrahydrofuran (10 mL)at −78° C. under nitrogen. The mixture was stirred at −78° C. for 20 minbefore the temperature was slowly raised to room temperature. Themixture was stirred at room temperature for 16 hr. The reaction mixturewas cooled to 0° C. and saturated aqueous NH₄Cl was added to quench thereaction. The reaction mixture was diluted with ethyl acetate and washedwith saturated aqueous NH₄Cl. The organic layer was dried with MgSO₄,filtered and concentrated. The crude material was purified on a silicagel cartridge (40 g) using an EtOAc/Hex gradient (0-100% EtOAc over 20CV) to afford(5R,7S)-7-((S)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(1.3 g, 3.50 mmol). HPLC retention time=1.09 min (condition A); LC/MSM⁺¹=372.5.

To a mixture of(5R,7S)-7-((S)-6-(5-methoxypentyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(680 mg, 1.830 mmol) in dioxane (20 mL) was added 1N NaOH (10 mL). Thereaction mixture was heated at 95° C. After stirring two days, thereaction mixture was cooled, diluted with ethyl acetate, and washed withsaturated NaCl. The organic layer was dried with MgSO₄, filtered andconcentrated to afford 450 mg of Example 672 as a white solid. HPLCretention time=7.1 min (condition L); LC/MS M⁺¹=346.

Alternative Preparation of Example 677

To a mixture of((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (440 mg, 0.966 mmol) and copper(I) bromide (277mg, 1.932 mmol) in THF (10 mL) at 0° C. was added(3-methoxybenzyl)magnesium chloride (35 ml, 8.75 mmol). The mixture wasstirred at 0° C. and was slowly raised to room temperature and stirredovernight.

The reaction mixture was cooled down to 0° C., 1 ml of water was addedand the mixture was mixed with EtOAc (80 ml) and water (20 ml). Theorganic phase was separated and washed with saturated NH₄Cl (3×30 ml)and brine (20 ml). The organic solution was dried over anhydrous sodiumsulfate and concentrated. Flash chromatography purification using ISCO(40 g silica gel column, gradient elution from 0 to 100% of EtOAc/hexanefor 13CV. Product containing fractions were isolated. Recovered(5R,7S)-7-((R)-6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(370 mg, 0.912 mmol). HPLC retention time=1.14 min (condition A); LC/MSM⁺¹=406.

To a mixture of(5R,7S)-7-((R)-6-(3-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(370 mg, 0.912 mmol) in dioxane (20 mL) was added 1N NaOH. The reactionmixture was heated at 95° C. overnight, cooled, diluted with ethylacetate, and then washed with saturated NaCl. The organic layer wasdried with MgSO₄, filtered and concentrated. The resulting solid wastriturated in MeCN and allowed to stir overnight. The mixture wasfiltered and the resulting solid material was dried in vacuo to afford255 mg of Example 677. HPLC retention time=7.73 min (condition L); LC/MSM⁺¹=380.

Alternative Preparation 1 of Example 679

A 1.0M THF solution of allylmagnesium bromide (8.78 mL, 8.78 mmol) wasadded to a stirred mixture of copper(I) bromide (126 mg, 0.878 mmol),((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (200 mg, 0.439 mmol) and anhydroustetrahydrofuran (5 mL) at −78° C. under nitrogen. The mixture wasstirred at −78° C. for 20 min before the temperature was raised to roomtemperature over 20 min. The mixture was stirred at room temperature for5 hr. Saturated aqueous NH₄Cl solution (5 mL) was added slowly to quenchthe reaction. Hexanes (7 mL) and water (1 mL) were added. The aqueouslayer was separated and extracted with ethyl acetate (2×3 mL). Thecombined organic solutions were dried over sodium sulfate andconcentrated under reduced pressure. Flash chromatography purificationusing ISCO (4 g silica gel column, gradient elution from 0 to 100% ofethyl acetate in hexanes) afforded(5R,7S)-7-((R)-6-(but-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(90 mg, 0.277 mmol). HPLC retention time=1.14 min (condition A); LC/MSM⁺¹=326.

To a clear solution of(5R,7S)-7-((R)-6-(but-3-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(560 mg, 1.721 mmol) in THF (30 mL) were sequentially added 50% NMO (403mg, 3.44 mmol) and osmium tetroxide in t-BuOH (0.647 mL, 0.052 mmol) atroom temperature. The solution was vigorously stirred at roomtemperature overnight. Sodium periodate (1472 mg, 6.88 mmol) in H₂O (15mL) was added. The mixture was stirred vigorously at room temperatureunder nitrogen for 30 min. The mixture was extracted with ethyl acetate(3×2 mL). The combined ethyl acetate extracts were dried (Na₂SO₄) andconcentrated. Flash chromatography purification using ISCO (40 g silicagel column, gradient elution from 20 to 100% of ethyl acetate inhexanes) afforded3-((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propanal(440 mg, 1.344 mmol). NMR was consistent with desired product. HPLCretention time=0.91 min (condition A); LC/MS M⁺¹=328.

To a stirred solution of3-((S)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)propanal(700 mg, 2.138 mmol), ethoxytrimethylsilane (1.670 mL, 10.69 mmol), andtriethylsilane (1.707 mL, 10.69 mmol) in nitromethane (5 mL) was addedferric chloride (34.7 mg, 0.214 mmol) at 0° C. under nitrogen. Themixture was stirred at 0° C. for 15 min and at room temperature for 12hours. The reaction mixture was diluted with ethyl acetate and washedwith saturated NaCl. The organic layer was dried with MgSO₄, filteredand concentrated. The crude material was purified on a silica gelcartridge (40 g) using an EtOAc/Hex gradient (0-100% EtOAc over 12 CV).Recovered(5R,7S)-7-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(660 mg, 1.846 mmol). HPLC retention time=1.06 min (condition A); LC/MSM⁺¹=356.

To a mixture of(5R,7S)-7-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(250 mg, 0.699 mmol) in dioxane (10 mL) was added 1N NaOH. The reactionmixture was heated at reflux for 48 hrs. The mixture was cooled, dilutedwith ethyl acetate, and washed with saturated NaCl. The organic layerwas dried with MgSO₄, filtered, and concentrated. Material was purifiedin batch by HPLC. HPLC conditions: Phenomenex Luna 5 micron C18 column(30×100 mm); MeCN (0.1% TFA)/water (0.1% TFA); 25%-100% gradient over 15minutes; 30 mL/min. Fractions with correct mass were isolated, pouredinto 1N NaOH, extracted with EtOAc (2 times), and then the pooled EtOAclayers were washed with 1 N NaOH one more time. The solution was driedand concentrated in vacuo to afford((1R,3S)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(186 mg, 0.554 mmol. HPLC retention time=7.08 min (condition L); LC/MSM⁺¹=332.

Alternative Preparation 2 of Example 679 Preparation of(5R,7S)-7-((R)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a solution of (R)-methyl6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carboxylate(40.9 g, 124 mmol) in THF (250 ml) was added a light suspension ofcalcium chloride (11.1 g, 100 mmol) in EtOH (250 ml) and the resultingsolution was cooled to 0° C. Sodium borohydride (7.7 g, 199 mmol) wasadded and the mixture was stirred at 0° C. for 2.0 h. At this time, themixture was allowed to warm up to room temperature and stirred for 36.5h. Then, the mixture was cooled to 0° C. and quenched with phosphatebuffer (1.5M KH₂PO₄+H₃PO₄ to pH 3, 500 mL, slow initial addition, gasevolution). The aqueous mixture was stirred at room temperature for 3.0h and then mixed with CH₂C2 (700 mL) in a separatory funnel. The pH ofthe aqueous layer was adjusted to 3 by addition of 6M HCl and thebiphasic mixture was shaken. The organic layer was collected and theaqueous phase was extracted with CH₂C2 (2×250 mL). The combined organiclayers were dried (Na₂SO₄) and concentrated. The resulting solid wastriturated with Et₂O and the suspension was filtered through a sinteredfunnel. The solid was rinsed with Et₂O, dried by suction, collected anddried under vacuum to afford(5R,7S)-7-((R)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(37.3 g) as a white solid. Analytical HPLC (Gemini method): RT=4.81 min,Area %: 100; LC/MS M⁺¹=302; ¹H NMR (400 MHz, CDCl₃) δ 7.04 (d, J=7.5 Hz,1H), 6.96 (m, 2H), 5.12 (s, 1H), 4.35 (d, J=8.4 Hz, 1H), 4.30 (d, J=8.4Hz, 1H), 3.66 (m, 2H), 3.05 (m, 1H), 2.86 (m, 3H), 2.51 (dd, J=16.3,10.7 Hz, 1H), 2.33 (dd, J=13.3, 7.3 Hz, 1H), 2.15 (m, 2H), 2.00 (m, 4H)1.84 (m, 1H), 1.52-1.36 (m, 2H).

Preparation of(R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carbaldehyde

A 2M oxalyl chloride/CH₂Cl₂ solution (25.0 ml, 50.0 mmol) was dilutedwith CH₂C2 (100 ml) and cooled to −78° C. while stirring. DMSO (7.1 ml,100 mmol) was slowly added to the resulting solution and the mixture wasstirred at −78° C. for 30 min. Then, a cloudy solution of(5R,7S)-7-((R)-6-(hydroxymethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(10.0 g, 33.3 mmol) in CH₂C2 (50 ml) and DMSO (8.0 ml) was added over a25 min period. After the addition was complete, stirring at −78° C. wascontinued for 30 min and after this time, triethylamine (14.0 ml, 100mmol) was added dropwise over a 15 min period. The reaction mixture wasstirred for 1.5 h at −78° C. and for 30 min while warming up to 0° C.The reaction was quenched at 0° C. with 1M KH₂PO₄ (150 mL). The biphasicmixture was shaken in a separatory funnel. The organic layer was washedwith water (150 mL) and saturated NaCl (150 mL), dried (Na₂SO₄) andconcentrated. Further drying under vacuum gave(R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carbaldehyde(10.6 g, 31.5 mmol) as a colorless solid. Analytical HPLC (Geminimethod): RT=5.77 min, Area %: 89; LC/MS M⁺¹=300; ¹H NMR (400 MHz, CDCl₃)9.81 (d, J=1.1 Hz, 1H), 7.12 (d, J=7.9 Hz, 1H), 7.00 (dd, J=7.9, 1.6 Hz,1H), 6.96 (s, 1H), 5.45 (s, 1H), 4.35 (d, J=8.4 Hz, 1H), 4.29 (d, J=8.4Hz, 1H), 3.11-2.79 (m, 5H), 2.73 (m, 1H), 2.33 (dd, J=13.4, 7.4 Hz, 1H),2.24 (m, 1H), 2.15 (m, 2H), 2.02-1.91 (m, 2H), 1.89-1.76 (m, 2H).

Preparation of (2-ethoxyethyl)triphenylphosphonium Bromide

To a 3-necked round bottom flask equipped with a mechanic stirrer wascharged with triphenylphosphine (41.7 g, 159 mmol) and toluene (550 mL).The solution was added 1-bromo-2-ethoxyethane (22.11 mL, 176 mmol) underN₂ at room temperature. The reaction mixture was heated to 95° C. for 18hrs. The solid was formed during the reaction. After 18 hours, thereaction mixture was cooled down to room temperature and stirred for 30minutes. The slurry was filtered, rinsed with toluene (2×100 ml) anddried under high vacuo to give (2-ethoxyethyl)triphenylphosphoniumbromide (60.1 g, 145 mmol) as an off-white solid. LC/MS M⁺¹=336.

Preparation of(5R,7S)-7-((R)-6-((Z)-3-ethoxyprop-1-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

1M Potassium tert-butoxide (48.6 mL, 48.6 mmol) was added over a 20 minperiod to a suspension of (2-ethoxyethyl)triphenylphosphonium bromide(20.8 g, 50.1 mmol) in THF (205 mL) at −78° C. and under Ar. After theaddition was complete, the mixture was stirred at −78° C. for 30 min andthen, a solution of(R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carbaldehyde(10.6 g, 31.5 mmol) in CH₂C2 (69 ml) was added dropwise over a 40 minperiod. The mixture was stirred for 17.5 h while slowly warming up to19° C. After cooling the reaction mixture to 0° C., 1M KH₂PO₄ (100 mL)was added. The resulting aqueous mixture was stirred at room temperaturefor 30 min and then extracted with EtOAc (300 mL). The organic extractwas washed with water (100 mL) and saturated NaCl (100 mL), dried(Na₂SO₄) and concentrated. The crude was purified by chromatography(SiO₂ 750 g gold RediSep column, 0 to 40% acetone/hexanes) to afford(5R,7S)-7-((R)-6-((Z)-3-ethoxyprop-1-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(8.93 g) as a white solid. Analytical HPLC (Gemini method): RT=8.65 min,Area %: 97; LC/MS M⁺¹=356; ¹H NMR (400 MHz, CDCl₃) 7.04 (d, J=7.7 Hz,1H), 6.97 (m, 2H), 5.59 (m, 2H), 5.10 (s, 1H), 4.35 (d, J=8.4 Hz, 1H),4.30 (d, J=8.4 Hz, 1H), 4.09 (d, J=5.2 Hz, 2H), 3.52 (q, J=7.0 Hz, 2H),3.05 (m, 1H), 2.85 (dd, J=8.3, 4.5 Hz, 2H), 2.83-2.71 (m, 2H), 2.57 (m,1H), 2.34 (dd, J=13.4, 7.4 Hz, 1H), 2.21-2.08 (m, 2H), 2.05-1.77 (m,2H), 1.61 (m, 1H), 1.24 (t, J=6.9 Hz, 3H).

Preparation of(5R,7S)-7-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

To a stirred solution of(5R,7S)-7-((R)-6-((Z)-3-ethoxyprop-1-en-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(10.8 g, 28.9 mmol) in THF (275 ml) was added platinum(IV) oxide (0.408g, 1.797 mmol). The resulting suspension was stirred under hydrogen (1atm, balloon) for 10.0 h. The suspension was filtered through Celite andthe filter cake was rinsed with CH₂C2 (200 mL) and MeOH (80 mL). Thefiltrate and rinses were combined and evaporated to give crude(5R,7S)-7-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(10.6 g) as a brownish solid. Analytical HPLC (Gemini method): RT=9.29min, Area %: 92.

The above crude material was filtered through a short pad of SiO₂(230-400 mesh) eluting with 4/1 to 7/3 CH₂Cl₂/EtOAc to obtain 10.0 g ofa material that contained a hydrogenolysis byproduct and an epimericimpurity not resolved by the above HPLC conditions. This later materialwas purified by SFC to afford(5R,7S)-7-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(7.3 g) as an off-white solid. Analytical HPLC (Gemini method): RT=9.39min, Area %: 99; LC/MS M⁺¹=358.

Preparation of((1R,3S)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

To a stirred solution of the(5R,7S)-7-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(20.5 g, 56.8 mmol) in 2-methyltetrahydrofuran (60.0 ml) and EtOH (120ml) was added a solution of lithium hydroxide (6.2 g, 254 mmol) in water(60.0 ml). The mixture was heated to 90° C. and stirred at thistemperature for 16.0 h. Then, the reaction mixture was cooled to roomtemperature and filtered through a sintered funnel. The white solidremaining in the sintered funnel was triturated and rinsed with CH₂C2(200 mL), then water (150 mL) and finally with additional CH₂C2 (200mL). The filtrate and rinses were combined and transferred to aseparatory funnel. The biphasic mixture was shaken and the layersseparated. The organic layer was collected, dried (Na₂SO₄) andconcentrated to give((1R,3S)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(18.8 g) as a brownish foam: HPLC (Gemini method): RT=4.74 min, Area %:99; LC/MS M⁺¹=332.

Preparation of((1R,3S)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,Hydrochloride

To a stirred solution of the((1R,3S)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(17.0 g, 50.8 mmol) in EtOH (115 ml) at room temperature was added 1.25MHCl/EtOH (50.0 ml, 62.4 mmol). The resulting solution was stirred for2.4 h and became a suspension. The solid that formed was collected byfiltration, rinsed with diethyl ether, then dissolved in MeOH andfiltered through the sintered funnel. The methanolic solution wasevaporated and dried under vacuum to obtain 13.9 g of a white solid. Thefiltered solution and ether rinses were combined and evaporated in vacuountil precipitation was observed. The solid that formed was isolated asdescribed above to give an additional 2.33 g of white solid. The abovesolids were combined to afford((1R,3S)-1-amino-3-((S)-6-(3-ethoxypropyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol,hydrochloride (16.2 g) as a white solid: HPLC (Gemini method): RT=4.77min, Area %: 99; LC/MS M⁺¹=332; ¹H NMR (500 MHz, CD₃OD) δ 7.00 (m, 3H),3.66 (d, J=11.6 Hz, 1H), 3.60 (t, J=11.6 Hz, 1H), 3.52 (q, J=7.0 Hz,2H), 3.49 (t, J=6.6 Hz, 2H), 3.11 (m, 1H), 2.90-2.71 (m, 3H), 2.40 (m,2H), 2.11 (m, 1H), 2.02-1.86 (m, 4H), 1.77-1.64 (m, 4H), 1.50-1.33 (m,3H), 1.21 (t, J=7.1 Hz, 3H).

Alternative Preparation-1 of Example 681

To a mixture of bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II) (23.97 mg, 0.034 mmol), cesium carbonate (331 mg,1.016 mmol), and(5R,7S)-7-((R)-6-ethynyl-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(100 mg, 0.339 mmol) in acetonitrile (10 mL) was added1-iodo-2-methoxybenzene (0.132 mL, 1.016 mmol). The reaction mixture wassparged with nitrogen for 5 minutes then heated at 70° C. overnight. Thereaction mixture was diluted with ethyl acetate and washed withsaturated NaCl. The organic layer was dried with MgSO₄, filtered andconcentrated. The crude material was purified on a silica gel cartridge(24 g) using an EtOAc/hexane gradient (0-100% EtOAc over 20 CV).Isolated fractions 14-15 were concentrated and dried in vacuo to afford(5R,7S)-7-((R)-6-((2-methoxyphenyl)ethynyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(70 mg, 0.174 mmol). HPLC retention time=1.07 min (condition A); LC/MSM⁺¹=402.

To a mixture of(5R,7S)-7-((R)-6-((2-methoxyphenyl)ethynyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(65 mg, 0.162 mmol) in MeOH (5 mL) was added Pearlman's Catalyst (5 mg,0.036 mmol). The reaction mixture was hydrogenated under a balloon of H₂for 1 hour. The mixture was filtered to remove catalyst. Next, 1 N NaOH(5 mL) was added and the mixture was heated at 95° C. overnight. Thereaction mixture was diluted with ethyl acetate and washed with H₂O. Theorganic layer was dried with MgSO₄, filtered and concentrated. Purifiedby HPLC. HPLC conditions: Phenomenex Luna 5 micron C18 column (30×100mm); MeCN (0.1% TFA)/water (0.1% TFA); 20%-100% gradient over 15minutes; 30 mL/min. Fractions with correct mass were pooled, then washedwith 1N NaOH and extracted with EtOAc. EtOAc layer was washed two moretimes and then back extracted the aqueous layer once. The organic layerwas dried with MgSO₄, filtered, concentrated, and freeze dried fromMeCN/water to afford((1R,3S)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(55 mg, 0.138 mmol). HPLC retention time=8.26 min (condition L); LC/MSM⁺¹=380.

Alternative Preparation-2 of Example 681

To a mixture of((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (1 g, 2.195 mmol) and potassium carbonate(0.910 g, 6.59 mmol) in DMF (10) was added 1-phenyl-1H-tetrazole-5-thiol(0.782 g, 4.39 mmol). The reaction mixture was heated at 80° C.overnight. The reaction mixture was diluted with ethyl acetate andwashed with saturated NaCl. The organic layer was dried with MgSO₄,filtered and concentrated. The crude material was purified on a silicagel cartridge (40 g) using an EtOAc/Hex gradient (0-100% EtOAc over 13CV) to afford(5R,7S)-7-((R)-6-(((1-phenyl-1H-tetrazol-5-yl)thio)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(0.94 g, 2.036 mmol). HPLC retention time=1.02 min (condition A); LC/MSM⁺¹=462.

To hydrogen peroxide (8.32 mL, 81 mmol) at 0° C. was added ammoniummolybdate tetrahydrate (0.503 g, 0.407 mmol). The resulting solution wasadded to a mixture of(5R,7S)-7-((R)-6-(((1-phenyl-1H-tetrazol-5-yl)thio)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(0.94 g, 2.036 mmol) in THF (15 mL) at 0° C. The reaction mixture wasstirred overnight. The reaction mixture was diluted with ethyl acetateand washed with saturated NaCl. The organic layer was dried with MgSO₄,filtered and concentrated to afford(5R,7S)-7-((R)-6-(((1-phenyl-1H-tetrazol-5-yl)sulfonyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(1 g, 2.026 mmol). HPLC retention time=0.96 min (condition A); LC/MSM⁺¹=494.

To a mixture of 2-methoxybenzaldehyde (497 mg, 3.65 mmol) and(5R,7S)-7-((R)-6-(((1-phenyl-1H-tetrazol-5-yl)sulfonyl)methyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(600 mg, 1.216 mmol) in THF was added KHMDS (4.86 mL, 4.86 mmol). Afterstirring at room temperature for 1 hour, the reaction was quenched withMeOH. The mixture was purified by HPLC. The crude material was purifiedon a silica gel cartridge (24 g) using an EtOAc/Hex gradient (0-100%EtOAc over 12 CV). Isolated fractions 18-20 were concentrated and driedin vacuo to afford(5R,7S)-7-((R)-6-((E)-2-methoxystyryl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(182 mg, 0.451 mmol). HPLC retention time=1.13 min (condition A); LC/MSM⁺¹=404.

To a mixture of(5R,7S)-7-((R)-6-((E)-2-methoxystyryl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(182 mg, 0.451 mmol) in MeOH (10 mL) was added Pearlman's Catalyst (5mg, 0.036 mmol). The reaction mixture was hydrogenated under a balloonof H₂ for 1 hour. The mixture was filtered to remove the catalyst, and 1N NaOH (5 mL) was added. The reaction mixture was heated at 95° C.overnight. The mixture was cooled, diluted with ethyl acetate, andwashed with H₂O. The organic layer was dried with MgSO₄, filtered andconcentrated. The solid material was triturated in MeCN and stirredovernight. Solids were collected by filtration and dried to afford((1R,3S)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(90 mg, 0.235 mmol). HPLC retention time=7.93 min (condition L); LC/MSM⁺¹=380.

Alternative Preparation-3 of Example 681

To a solution of((R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)methyl4-methylbenzenesulfonate (700 mg, 1.537 mmol) and copper(I)bromide-dimethyl sulfide complex (948 mg, 4.61 mmol) in Et₂O (50 mL) wasadded (2-methoxybenzyl)magnesium chloride (58 ml, 14.50 mmol) at roomtemperature. The reaction mixture was stirred for 16 h. The reactionmixture was diluted with saturated NH₃Cl and water, and extracted withEtOAc. The organic layer was collected, dried over Na₂SO₄, concentratedto give 580 mg of desired product, M+H=406. This material was dissolvedin dioxane (10 mL) and 1N NaOH was added (10 mL). The mixture was heatedat 100° C. overnight. The mixture was cooled, diluted with water, andextracted with EtOAc (2×). The combined organic layer was washed withsaturated NaCl, then dried over MgSO₄, and concentrated on therotavapor. The solid material was triturated in MeCN (10 mL) and thenstirred for several hours. The solid material was collected byfiltration and dried in vacuo to give((1R,3S)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol(350 mg, 0.876 mmol). MS (m+1)=380; HPLC Peak RT=9.06 min (Condition L)Purity=97%.

Alternative Preparation-4 of Example 681 Preparation of5-((2-methoxybenzyl)thio)-1-phenyl-1H-tetrazole

Sodium carbonate (135 g, 1277 mmol) was added portionwise to a solutionof 1-(chloromethyl)-2-methoxybenzene (200 g, 639 mmol) and1-phenyl-1H-tetrazole-5-thiol (125 g, 702 mmol) in anhydrous DMF (639ml). The reaction mixture was allowed to stir at room temperature undera nitrogen atmosphere for 2 days before diluting with water (1000 ml)and extracting with ethyl acetate (3×300 ml). The combined organics werethen washed with water (500 mL), brine (500 mL) and then dried (MgSO₄).The solvent was evaporated in vacuo and the crude purified by columnchromatography using ethyl acetate hexane as eluent to give5-((2-methoxybenzyl)thio)-1-phenyl-1H-tetrazole (167 g, 88%) as a whitesolid. HPLC retention time (Sunfire C18 Sum 4.6×50 (4 min grad.) SolventA=10% MeOH-90% H₂O-0.2% H₃PO₄; Solvent B=90% MeOH-10% H₂O-0.2%H₃PO₄)=3.34 min. ¹H NMR (400 MHz, DMSO-d₆) δ 7.73-7.48 (m, 5H), 7.37(dd, J=7.5, 1.8 Hz, 1H), 7.29 (td, J=7.8, 1.8 Hz, 1H), 6.99 (d, J=8.1Hz, 1H), 6.88 (td, J=7.5, 0.9 Hz, 1H), 4.53 (s, 2H), 3.76 (s, 3H).

5-((2-methoxybenzyl)sulfonyl)-1-phenyl-1H-tetrazole

To a 500 mL, 3 neck round bottom flask was added hydrogen peroxide(0.274 L, 2681 mmol). The contents of the flask were cooled to 0-5° C.Ammonium molybdate tetrahydrate (66.3 g, 53.6 mmol) was addedportionwise over 10 minutes while maintaining the temperature below 5°C. To a separate 5L, 3 neck round bottom flask with a mechanical stirrerwas added 5-((2-methoxybenzyl)thio)-1-phenyl-1H-tetrazole (80 g, 268mmol) in acetonitrile (2L). The peroxide solution was added slowly whilemaintaining the temperature below 30° C. during the addition. A yellowsuspension formed. The reaction mixture was stirred at room temperaturefor 18 h. The reaction mixture was cooled with an ice bath to 5° C. anddiluted with water (2.7 L) and stirred for 1 h. The suspension wasfiltered and washed with water and dried by vacuum suction to give amixture of sulfone and sulfoxide (˜80 g) which was re-subjected to theoxidation conditions described above to give the crude product (˜80 g)which was then purified by column chromatography using ethyl acetatehexane as eluent to give5-((2-methoxybenzyl)sulfonyl)-1-phenyl-1H-tetrazole (71 g, 215 mmol) asa white crystalline solid. HPLC retention time (BEH C18 2.1×50 mm 1.7um, 2 min grad., Solvent Name A: 100% H₂O w/0.05% TFA; Solvent Name B:100% ACN w/0.05% TFA) 0.91 min. ¹H NMR (400 MHz, CHLOROFORM-d) δ7.62-7.54 (m, 1H), 7.54-7.45 (m, 2H), 7.41 (ddd, J=8.3, 7.5, 1.8 Hz,1H), 7.37-7.30 (m, 3H), 6.96 (td, J=7.5, 1.0 Hz, 1H), 6.91 (d, J=8.4 Hz,1H), 5.02 (s, 2H), 3.74 (s, 3H).

(5R,7S)-7-((R)-6-((E)-2-methoxystyryl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

LiHMDS (46.8 ml, 70.1 mmol) was added dropwise to a solution of5-((2-methoxybenzyl)sulfonyl)-1-phenyl-1H-tetrazole (23.17 g, 70.1 mmol)in anhydrous THE (84 ml) and DMF (55.9 ml) at −78° C. under a nitrogenatmosphere. The addition took ˜5 min and the temperature of the reactionmixture did not rise above −60° C. The resulting solution was coloredorange. The reaction mixture was allowed to stir at −78° C. for 30 minbefore the dropwise addition of(R)-6-((5R,7S)-2-oxo-3-oxa-1-azaspiro[4.4]nonan-7-yl)-1,2,3,4-tetrahydronaphthalene-2-carbaldehyde(10 g, 33.4 mmol) in anhydrous DMF (39.9 ml) [as 14 mL+6 mL washing].The temperature did not rise above −70° C. during the addition. Thereaction mixture was allowed to warm slowly to room temperatureovernight. HPLC indicated desired product. The reaction mixture wascooled to −78° C. before the reaction was quenched with water (20 mL).The mixture was allowed to warm to room temperature. The reactionmixture was partitioned between water and ethyl acetate, and the aqueouslayer was extracted with ethyl acetate (2×). The combined organics werethen washed with water, brine, and dried (MgSO₄). The evaporated organiclayer was then purified by column chromatography using ethylacetate:hexane as eluent to give(5R,7S)-7-((R)-6-((E)-2-methoxystyryl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(12.1 g, 90%) as a white solid. HPLC retention time (BEH C18 2.1×50 mm1.7 um, 2 min grad., Solvent Name A: 100% H₂O w/0.05% TFA; Solvent NameB: 100% ACN w/0.05% TFA): 1.27 and 1.28 min, as a 1:2 mixture of doublebond isomers. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.48 (dd, J=7.7, 1.5 Hz,1H), 7.33-7.18 (m, 5H), 7.13-7.02 (m, 2H), 7.01-6.87 (m, 6H), 6.83 (d,J=16.1 Hz, 1H), 6.56 (d, J=11.7 Hz, 1H), 6.29 (dd, J=16.1, 6.8 Hz, 1H),5.69 (dd, J=11.7, 9.9 Hz, 1H), 5.15 (br. s., 1H), 4.40-4.25 (m, 3H),3.87 (d, J=4.0 Hz, 5H), 3.13-2.78 (m, 7H), 2.76-2.58 (m, 3H), 2.41-2.27(m, 2H), 2.22-2.05 (m, 4H), 2.04-1.91 (m, 4H), 1.89-1.75 (m, 2H),1.74-1.60 (m, 2H), 1.57 (s, 4H).

(5R,7S)-7-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

(5R,7S)-7-((R)-6-((E)-2-methoxystyryl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(24 g, 59.5 mmol) was dissolved in dichloromethane (297 ml) and MeOH(297 ml). Nitrogen gas was bubbled through the solution for about 10minutes. Next, Pd/C (6.33 g, 5.95 mmol) was added in one portion. Ahydrogen filled balloon was placed on the reaction flask beforeevacuating the flask for ˜2 minutes. Hydrogen was then introduced andthis process was repeated 2 more times. After stirring at roomtemperature overnight, HPLC indicated conversion to the desired productbut with significant starting material remained. The reaction mixturewas filtered through Celite, washed with 1:1 MeOH:DCM and the filtratewas evaporated in vacuo. The residue was set up as described above andresubjected to the hydrogenation overnight. HPLC still shows startingmaterial remaining. A third hydrogenation gave complete conversion tothe desired product. The reaction mixture was filtered through Celiteand evaporated. SFC was performed to remove a minor undesired isomer andgave(5R,7S)-7-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(24.67 g, 60.8 mmol) as a white solid. HPLC retention time (Sunfire C185 um 4.6×50 (4 min grad.) Solvent A=10% MeOH-90% H₂O-0.2% H₃PO₄; SolventB=90% MeOH-10% H₂O-0.2% H₃PO₄)=4.15 min.((1R,3S)-1-amino-3-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)cyclopentyl)methanol

LiOH (21.91 g, 915 mmol) was added in one portion to a solution of(5R,7S)-7-((S)-6-(2-methoxyphenethyl)-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(53 g, 131 mmol) in dioxane (523 ml) and H₂O (131 ml). The reactionmixture was allowed to reflux under a nitrogen atmosphere for 72 hours.The reaction mixture was allowed to cool to room temperature beforediluting with water (300 ml) and extracting with ethyl acetate (3×300ml). The combined organics were then dried (MgSO₄) and evaporated invacuo. The residue was recrystallized from IPA to give a white solid (27g). The remainder of the material formed a chelate with the dryingagent. This material was treated with hot ethanol and the resultingslurry filtered to remove the drying agent. The filtrate was evaporatedin vacuo and taken up in ethyl acetate (300 ml) before washing with 1Nsodium hydroxide solution (200 ml). The organic layer was dried (Na₂SO₄)and evaporated to give a residue which was recrystallized from IPA togive a white solid (13 g). The remainder of the material (filtrates fromthe above two manipulations) was purified by SFC to give 10 g of a whitesolid. HPLC retention time (Sunfire C18 Sum 4.6×50 (4 min grad.) SolventA=10% MeOH-90% H₂O-0.2% H₃PO₄; Solvent B=90% MeOH-10% H₂O-0.2%H₃PO₄)=3.21 min.

Comparative Compound 703(1R,3R)-1-Amino-3-(6-(pentyloxy)naphthalen-2-yl)cyclopentyl)methanol

Comparative Compound 703 was disclosed in WO 2008/079382, Example Q.1.

Intermediate 703A:(5R,7R)-7-(6-(Pentyloxy)naphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one

A mixture of 1-pentanol (6.13 mL, 56.4 mmol), p-toluenesulfonic acidmonohydrate (4.60 mg, 0.024 mmol), and trimethoxymethane (0.353 mL, 3.22mmol) was stirred at 100° C. for 3 hr with a slow air stream flowingover the mixture to remove methanol and some pentanol. The obtainedresidual liquid was mixed with(5R,7R)-7-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(Int. 7, 230 mg, 0.806 mmol) and stirred at 100° C. under nitrogen for2.5 hr. The solution was allowed to cool down to room temperature beforepalladium on carbon (172 mg, 0.081 mmol) was added, followed by ethylacetate (4 mL). The mixture was left to stir under a balloon-pressure ofhydrogen at room temperature overnight. The resulting mixtures werefiltered through a membrane filter and the filtrate was concentrated.Flash chromatography purification (24 g silica gel column, 0% to 70%ethyl acetate in hexanes) afforded 180 mg of material that requiredadditional purification. Supercritical Fluid Chromatographic separationafforded a major fraction by UV analysis identified as(5R,7R)-7-(6-(pentyloxy)naphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(36 mg) as a solid. Instrument: Thar 350 Thar Analytical SFC-MS;Conditions: Analytical Conditions: Analytical Column: AD-H (0.46×25 cm,5 μm); BPR pressure: 100 bars; Temperature: 45° C.; Flow rate: 3.0mL/min; Mobile Phase: CO₂/MeOH (70/30); Detector Wavelength: UV 200-400nm. Preparative Conditions: Preparative Column: AD-H (3×25 cm, 5 μm);BPR pressure: 100 bars; Temperature: 35° C.; Flow rate: 120 mL/min;Mobile Phase: CO₂/MeOH (70/30); Detector Wavelength: 220 nm; Separationprogram: Stack injection; Injection: 2.5 mL with cycle time 480 sec.(Analytical SFC ret. time=11.68 min, purity >99.5%) HPLC retentiontime=1.11 min (Condition G); LC/MS M⁺¹=354. ¹H NMR (400 MHz,chloroform-d) δ 7.68 (d, J=8.4 Hz, 2H), 7.55 (s, 1H), 7.30 (s, 1H),7.21-7.04 (m, 2H), 6.48 (br. s., 1H), 4.50-4.28 (m, 2H), 4.07 (t, J=6.6Hz, 2H), 3.49-3.31 (m, 1H), 2.46 (dd, J=13.3, 7.6 Hz, 1H), 2.39-2.24 (m,1H), 2.24-2.12 (m, 1H), 2.12-2.00 (m, 1H), 2.00-1.90 (m, 1H), 1.90-1.76(m, 3H), 1.58-1.30 (m, 4H), 0.96 (t, J=7.0 Hz, 3H).

Comparative Compound 703

To a solution of(5R,7R)-7-(6-(pentyloxy)naphthalen-2-yl)-3-oxa-1-azaspiro[4.4]nonan-2-one(36 mg, 0.102 mmol) in dioxane (2 mL) and water (0.8 mL) was added LiOH(36.6 mg, 1.528 mmol). The solution was heated to 90° C. and allowed tostir for 15 hours. The reaction mixture was cooled to room temperatureand was poured into ethyl acetate and washed with water. The crudematerial was then purified on reverse phase HPLC [Column: Luna Axia30*100 mm; Gradient time: 10 min; Flow rate=40 ml/min; Solvent A=10%MeOH-90% Water-0.1% TFA; Solvent B=90% MeOH-10% water-0.1% TFA; Start %B=20; Final % B=100]. The product containing fractions were collectedand dried under high vacuum to provide((1R,3R)-1-amino-3-(6-(pentyloxy)naphthalen-2-yl)cyclopentyl)methanol,TFA (31 mg) as a solid. HPLC retention time=0.90 min (Condition G);LC/MS M⁺¹=328. ¹H NMR (400 MHz, methanol-d₄) δ 7.75-7.66 (m, 2H),7.66-7.59 (m, 1H), 7.40-7.33 (m, 1H), 7.17 (d, J=2.6 Hz, 1H), 7.14-7.08(m, 1H), 4.07 (t, J=6.5 Hz, 2H), 3.74-3.60 (m, 2H), 3.59-3.41 (m, 1H),2.39-2.22 (m, 3H), 2.04-1.80 (m, 5H), 1.55-1.34 (m, 4H), 1.01-0.89 (m,3H).

Biological Assays

The compounds of Formulas (Ia), (IIa), (IIIa), (IVa), and (Va) or saltsthereof engage their biological targets (e.g. S1P1) after bioactivationthrough phosphorylation of the alcohol to provide an active phosphateester compound of Formulas (Ib), (IIb), (IIIb), (IVb), and (Vb), orsalts thereof. In vitro characterization of biological activity of theexamples was conducted on synthetically prepared samples of thephosphorylated compounds.

S1P₁ Binding Assay:

Membranes were prepared from CHO cells expressing human S1P₁. Cellspellets (1×10⁹ cells/pellet) were suspended in buffer containing 20 mMHEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), pH 7.5, 50mM NaCl, 2 mM EDTA (Ethylenediaminetetraacetic acid) and ProteaseInhibitor cocktail (Roche), and disrupted on ice using the Polytronhomogenizer. The homogenate was centrifuged at 20,000 rpm (48,000 g) andthe supernatant was discarded. The membrane pellets were resuspended inbuffer containing 50 mM HEPES, pH 7.5, 100 mM NaCl, 1 mM MgCl₂, 2 mMEDTA and stored in aliquots at −80° C. after protein concentrationdetermination.

Membranes (2 μg/well) and 0.03 nM final concentration of ³³P-S1P ligand(1 mCi/ml, Perkin elmer or American Radiolabeled Chemicals) diluted inassay buffer (50 mM HEPES, pH7.4, 5 mM MgCl2, 1 mM CaCl₂), 0.5% fattyacid free BSA (bovine serum albumin), 1 mM NaF) were added to thecompound plates (384 Falcon v-bottom plate (0.5 l/well in a 11 point,3-fold dilution). Binding was performed for 45 minutes at roomtemperature, terminated by collecting the membranes onto 384-wellMillipore FB filter plates, and radioactivity was measured by TOPCOUNT®.The competition data of the test compounds over a range ofconcentrations was plotted as percentage inhibition of radioligandspecific binding. The IC₅₀ is defined as the concentration of competingligand needed to reduce specific binding by 50%. The IC₅₀ for Example689 was determined to be 1.7 nM.

TABLE A S1P₁ Ex. Binding No. IC₅₀ (nM) 689 1.7 690 3.0 692 2.3 693 5.2695 118.9 698 69.4 699 183.4 701 48.7 702 151.1Receptor [³⁵S] GTPγS Binding Assays: (S1P₁ GTPγS/S1P₃ GTPγS)

Compounds were loaded in a 384 Falcon v-bottom plate (0.5 l/well in a 11point, 3-fold dilution). Membranes prepared from S1P₁/CHO cells orEDG3-Ga15-bla HEK293T cells (EDG3 equivalent S1P₃) were added to thecompound plate (40 l/well, final protein 3 μg/well) with MULTIDROP®.[³⁵S]GTP (1250 Ci/mmol, Perkin Elmer) was diluted in assay buffer: 20 mMHEPES, pH7.5, 10 mM MgCl2, 150 mM NaCl, 1 mM EGTA (ethylene glycoltetraacetic acid), 1 mM DTT (Dithiothreitol), 10 μM GDP, 0.1% fatty acidfree BSA, and 10 μg/ml Saponin to 0.4 nM. 40 μl of the [³⁵S] GTPsolution was added to the compound plate with a final concentration of0.2 nM. The reaction was kept at room temperature for 45 min. At the endof incubation, all the mixtures in the compound plate were transferredto Millipore 384-well FB filter plates via the VELOCITY11® Vprep liquidhandler. The filter plate was washed with water 4 times by using themanifold Embla plate washer and dried at 60° C. for 45 min. MicroScint20 scintillation fluid (30 l) was added to each well for counting on thePackard TOPCOUNT®. EC₅₀ is defined as the agonist concentration thatcorresponds to 50% of the Ymax (maximal response) obtained for eachindividual compound tested. The EC₅₀ for Example 689 was determined tobe 5.7 nM in the assay utilizing membranes prepared from S1P₁/CHO cells.The EC₅₀ for Example 689 was determined to be >2000 nM in the assayutilizing membranes prepared from EDG3-Ga15-bla HEK293T cells.

A smaller value for GTPγS S1P₁ EC₅₀ value indicated greater activity forthe compound in the GTPγS S1P₁ binding assay. A larger value for theGTPγS S1P3 EC₅₀ value indicated less activity in the GTPγS S1P3 bindingassay. Example 689, which is the phosphate ester of Example 672,possessed activity as an agonist of S1P₁ and is selective over S1P3.Example 697, which is the phosphate ester of Example 681, possessedactivity as an agonist of S1P₁ and is selective over S1P₃. Thus thecompounds of the present invention may be used in treating, preventing,or curing various S1P₁ receptor-related conditions while reducing orminimizing the side effects due to S1P₃ activity. The selectivity of thecompounds of the present invention indicate their potential use intreating, preventing, or curing autoimmune and inflammatory diseasessuch as multiple sclerosis, rheumatoid arthritis, inflammatory boweldisease, lupus, psoriasis, or vascular diseases, while reducing orminimizing possible side effects due to S1P₃ activity. Other potentialuses of the compounds of the present invention include minimizing orreducing rejection of transplanted organs, while reducing or minimizingside effects due to S1P₃ activity.

TABLE B Ex. GTPγS S1P₁ GTPγS S1P₃ No. EC₅₀ (nM) EC₅₀ (nM) 689 5.7 >2000690 21.4 >625 692 4.4 >625 693 19.2 >4162 694 38.6 >1250 695 49.0 >625696 1.2 >1000 697 0.6 >1000 698 8.9 >625 699 82.6 >625 701 133.4 >1250702 120.2 >625hS1P₁ ERK Phosphorylation (S1P₁ pERK)

hS1P₁/CHO cells were plated into BD Amine 384-well plates the day beforethe assay. On the day of the assay, growth medium was removed andreplaced with serum-free medium (Ham's F-12 Invitrogen) and incubatedfor 2 hours. Test compounds pre-diluted in HBSS (Gibco)/20 mM HEPES(Gibco) were transferred to the cell plates and incubated for 7 minutesat 37° C. Cells were lysed in lysis buffer (Perkin Elmer) andphospho-ERK was measured using the SureFire pERK kit (Perkin Elmer) asdescribed by the manufacturer. Data was plotted as percentage activationof the test compound relative to the efficacy of 10 μM S1P. The EC₅₀ isdefined as the concentration of test compound which produces 50% of themaximal response and was quantified using the 4 parameter logisticequation to fit the data. Data for phosphate examples in this assay areshown in Table I.

Blood Lymphocyte Reduction (BLR) Assay in Rodent:

Lewis rats or BALB/c mice were dosed orally with vehicle alone(polyethylene glycol 300, “PEG300”) or with test compounds. Compoundswere dosed as a solution or suspension in the vehicle, adjusted toreflect the free amount of test article in the event that salt forms areutilized. Blood was drawn at 24 hr and blood lymphocyte counts weredetermined on an ADVIA 120 Hematology Analyzer (Siemens HealthcareDiagnostics). The results were measured as a reduction in the percentageof circulating lymphocytes as compared to the vehicle treated group atthe time of measurement. The results represent the average results ofall animals within each treatment group (n=2-4). The results of theBlood Lymphocyte Reduction assay (BLR) in rat described hereinabove areshown in Table C.

The stereochemical orientation of the compounds in the present inventionwas found to influence the activity in the rodent BLR assay. Forexample, the diastereomeric set of compounds Examples 672, 673, 674, and675 were evaluated at the same dosage level of 0.1 mg/kg and theresulting lymphocyte reduction at 24 hours post-dose was found to rangefrom 30% for Example 674 to 63% for Example 675. The diastereomericcompounds Examples 678 and 679 were each evaluated at 0.1 mg/kg and theresulting lymphocyte reduction was found to be 16% and 65% respectively.Likewise, diastereomeric compounds Examples 681 and 682 were eachevaluated at 0.1 mg/kg and the resulting lymphocyte reduction was foundto be 53% and 17%, respectively.

TABLE C-1 Percent reduction vs. Ex. Dosage control at 24 hr post- No.(mg/kg) dose 672 0.1 47% 673 0.1 62% 674 0.1 30% 0.3 73% 675 0.1 63% 6760.1 63% 677 0.05 49% 678 0.1 16% 0.3 53% 679 0.1 65% 681 0.1 53% 682 0.117% 2.0 83% 684 0.05 72% 685 0.1 55%

The compounds of the present invention, as exemplified by Examples 679,681, and 684, have been compared to Comparative Compound 703, disclosedin WO 2008/079382, and have been found to be advantageous. As shown inTable C-2, Examples 679, 681, and 684 administered to mice at a dose of0.5 mg/kg, showed lymphocyte reductions of 59%, 85%, and 79%,respectively, at 24 hours post dose in this study. In comparison,Comparative Compound 703 administered a dose of 1.0 mg/kg, showed alymphocyte reduction of 52% at 24 hours post dose.

TABLE C-2 Mouse Blood Lymphocyte Reduction Assay Example or Dosage at 24hr post-dose Compound No (mg/kg) Percent reduction vs. control 679 0.559% 681 0.5 85% 684 0.5 79% 703 1.0 52%

The compounds of the present invention possess activity as agonists ofthe S1P₁ receptor, leading to the reduction of circulating bloodlymphocytes, and thus may be used in treating, preventing, or curingvarious S1P₁ receptor-related conditions. The surprising selectivity ofthe compounds of the present invention indicate their potential use intreating, preventing, or curing autoimmune and inflammatory diseasessuch as multiple sclerosis, rheumatoid arthritis, inflammatory boweldiseases, lupus, psoriasis, or vascular diseases. Other potential usesof the compounds of the present invention include minimizing or reducingrejection of transplanted organs.

Rat Adjuvant Induced Arthritis Assay (AA)

The rat adjuvant-induced arthritis model is an animal model for humanrheumatoid arthritis.

Male Lewis rats (150-175 g; Harlan, n=8 treatment group) were immunizedat the base of the tail with 100 μl of 10 mg/ml freshly groundMycobacterium butyricum (Difco Laboratories) in incomplete Freund'sadjuvant (sigma). Animals were dosed once daily with the test article(as a solution or suspension in the vehicle) or vehicle alone(polyethylene glycol 300, “PEG300”) starting from the day ofimmunization. The volumes of their hind paws were measured in a waterdisplacement plethysmometer (Ugo Basile, Italy). The baseline pawmeasurements were taken before onset of the disease (between day 7 today 10). The paw measurements were then taken three times a week untilthe end of the study on day 20 to 21. All procedures involving animalswere reviewed and approved by the Institutional Animal Care UseCommittee.

Example 672 of the present invention was tested in rat AA assaydescribed hereinabove and the results are shown in Table D. Thecompounds of this invention, as exemplified by Example 672, in thereported test, showed inhibition of disease progression as measured byreduced paw swelling in the Lewis rat using a prophylactic oral dosingregimen.

TABLE D Group paw swelling (mL) on day 20 Vehicle Mean 2.63  SEM* 0.14Example 672 Mean 2.60 (0.1 mg/kg) SEM 0.36 Example 672 Mean 1.46 (0.3mg/kg) SEM 0.34 Example 672 Mean 0.17 (1.0 mg/kg) SEM 0.08 *SEM:standard error of the mean

Example 679 was tested in the rat AA assay described hereinabove and theresults are shown in Table E. The compounds of this invention, asexemplified by Example 679 in the reported test, showed inhibition ofdisease progression as measured by reduced paw swelling in the Lewis ratusing a prophylactic oral dosing regimen.

TABLE E Group paw swelling (mL) on day 22 Vehicle Mean 1.62 SEM 0.24Example 679 Mean 1.55  (0.1 mg/kg) SEM 0.22 Example 679 Mean 0.36  (0.5mg/kg) SEM 0.19 Example 679 Mean 0.00 (2.50 mg/kg) SEM 0.05Mouse Experimental Autoimmune Encephalomyelitis Assay (EAE)

Mice (C57BL/6 female, 6-8 weeks of age, Charles River, n=10 treatmentgroup) were immunized subcutaneously with 150 μg MOG₃₅₋₅₅ emulsified 1:1with incomplete Freund's adjuvant (sigma) supplemented with 150 μgMycobacterium tuberculosis H37RA (Difco Laboratories). 400 ng ofpertussis toxin (CalBiochem) was injected intraperitoneally on the dayof immunization and 2 day later. Clinical scoring and body weight weretaken 3 times per week. Clinical scoring system: 0.5: partial tailweakness; 1: limp tail or waddling gait with tail tonicity; 1.5:waddling gait with partial tail weakness; 2: waddling gait with limptail (ataxia); 2.5: ataxia with partial limb paralysis; 3: fullparalysis of one limb; 3.5: full paralysis of one limbs with partialparalysis of a second limb; 4: full paralysis of two limbs; 4.5:moribund; 5: death. Mean clinical score was calculated by averaging thescores of all mice in each group. All procedures involving animals werereviewed and approved by the Institutional Animal Care Use Committee.

Example 681 of the present invention was tested in the mouse EAE assaydescribed herein above and the results are shown in Table F. Thecompounds of this invention, as exemplified by Example 681, in thereported test, showed inhibition of disease progression as measured byclinical scores in C57Bl/6 mice using a prophylactic oral dosingregimen.

TABLE F Group Clinical scores on day 22 Vehicle Mean 4.1 SEM 0.03Example 681 Mean 3.1 (0.1 mg/kg) SEM 0.16 Example 681 Mean 1.1 (0.5mg/kg) SEM 0.1 Example 681 Mean 0.8   (2 mg/kg) SEM 0.12

Example 679 of the present invention was tested in the mouse EAE assaydescribed herein above and the results are shown in Table G. Thecompound of this invention, as exemplified by Example 679, in thereported test, showed inhibition of disease progression as measured byclinical scores in C57Bl/6 mice using a prophylactic oral dosingregimen.

TABLE G Group Clinical scores on day 21 Vehicle Mean 4.1 SEM 0.03Example 679 Mean 2.9 (0.6 mg/kg) SEM 0.14 Example 679 Mean 1.8   (3mg/kg) SEM 0.14 Example 679 Mean 1.3  (15 mg/kg) SEM 0.08

In the mouse experimental autoimmune encephalomyelitis (EAE) model, ananimal model for multiple sclerosis, Examples 679 and 681 inhibitdisease progression as determined by the clinical scores in C57Bl/6 miceusing a prophylactic oral dosing regimen.

Rat Experimental Autoimmune Encephalomyelitis (EAE)

Female Lewis rats (150-200 g; Harlan) were immunized at the base of thetail with 0.1 ml of a complete Freund's adjuvant emulsion containing 0.5mg/mL guinea pig myelin basic protein (Genemed Synthesis) and 2 mg/mLMycobacterium butyricum (Difco). Beginning on Day 7, rats (n=11/group)were scored individually at least 3×/wk according to the followingscheme:

Score Clinical presentation 0.25 paralysis in the distal tail 0.5 limptail 1 ataxia (waddling gait with limp tail) 2 hind-leg paresis 3 fullhind-leg paralysis 4 Moribund 5 Death

Average clinical scores were calculated for each treatment group on eachday of assessment.

Example 679 of the present invention was tested in rat EAE assaydescribed hereinabove and the results are shown in Table H. The compoundof this invention, as exemplified by Example 679, in the reported test,showed inhibition of disease progression as measured by reduced clinicalscores in the Lewis rat using a prophylactic oral dosing regimen.

TABLE H Group Clinical Score on Day 11 Vehicle Mean 2.18 SEM 0.07Example 679 Mean 0.36 (0.3 mg/kg) SEM 0.08 Example 679 Mean 0.09 (1.0mg/kg) SEM 0.01 Example 679 Mean 0.02 (3.0 mg/kg) SEM 0.01MRL/Lpr Lupus Model:

MRL/lpr is a spontaneous model of lupus. Male MRL/lpr mice (JacksonLaboratory) at the age of 12-14 weeks were enrolled for the study(N=12). Mice were dosed p.o. daily with vehicle (18.4% (w/v)hydroxypropyl-b-cyclodextrin in 13.8 mM citric acid) or with Example 681at 0.06, 0.3, 1.5 mg/kg. Mice were bled every other week for anti-dsDNAantibodies measured by ELISA using pooled serum from diseased MRL/lprmice as a positive comparator in each assay. The data were expressed inarbitrary units as a ratio of the titer of the test serum to the titerof the pooled MRL/lpr immune serum.

At the end of study, one kidney was collected into 10% neutral bufferedformalin and ZincTris fixatives. Fixed tissues were processed intoparaffin blocks, sectioned at 3p m, and stained with H&E or PASH. Kidneysections were graded using following criteria: Glomerular Damage: 1.Mesangial matrix thickening, cell proliferation, 2. Crescent formation,cellular deposits/casts in Bowman's space, 3. Cellular infiltration,mononuclear cells in glomerular tufts, 4. Fibrosis of Bowman's capsule.Tubular damage: 1. Infiltration of mononuclear cells, 2. Severity oftubular damage, 3. Protein casts. Tubulo-interstitial damage: 1.Fibrosis, 2. Infiltration of mononuclear cells. Each subcategory wasassigned a score from 0-4, with the scores for glomerular indicesrepresenting the mean from 20 glomeruli per kidney. The total score foreach mouse was the sum of the above 9 subcategories, with the highestpossible score=36.

Example 681 was tested in NHI/lpr lupus model described herein above andthe results are shown in Table I-1 for anti-dsDNA antibody titers andTable I-2 for kidney histological analysis. Compounds of this invention,as exemplified by Example 681, in the reported test, showed inhibitionof disease progression as measured by anti-dsDNA titers and kidneyhistology.

TABLE I-1 Group Anti-dsDNA antibody titers (23 weeks of age) VehicleMean 3.328 SEM 0.660 Example 681 Mean 1.861 (0.06 mg/kg) SEM 0.581Example 681 Mean 0.978  (0.3 mg/kg) SEM 0.179 Example 681 Mean 1.023 (1.5 mg/kg) SEM 0.179

TABLE I-2 Kidney Histology (nephritis) 23 Group weeks of age VehicleMean 20.13 SEM 3.182 Example 681 Mean 10.13 (0.06 mg/kg) SEM 5.793Example 681 Mean 10.00  (0.3 mg/kg) SEM 3.295 Example 681 Mean 12.63 (1.5 mg/kg) SEM 4.719

In Table J, in vitro activity data determined by one or more of thefollowing assays: S1P₁ binding assay, receptor [³⁵S] GTPγS bindingassays (S1P₁ GTPγS/S1P₃ GTPγS), or hS1P₁ ERK Phosphorylation assay (S1P₁pERK) are shown for representative phosphate examples of this invention.

TABLE J S1P₃ S1P₁ Binding S1P₁ GTPγS S1P₁ pERK GTPγS A = (<10 nM) A =(<10 nM) A = (<10 nM) I = (>625 B = (10-100 nM) B = (10-100 nM) B =(10-100 nM) nM) C = (100- C = (100- C = (100- II = (>100 1000 nM) 1000nM) 1000 nM) nM) Ex. D = (1000- D = (1000- D = (1000- III = (>50 No.10000 nM) 10000 nM) 10000 nM) nM) 412 A A A I 414 A B D I 415 B B B I416 D — — I 417 B C A I 419 — — B I 420 B B D I 421 A A A I 423 C C B I424 B B A I 425 A A A I 426 B A A I 427 B A A I 428 A A B I 429 A A B I430 B B A I 431 B C A I 432 A A B II 433 B C A I 434 A B A I 435 B — — I436 A B A I 437 A A A I 438 A B A I 439 A B A I 440 A A A I 441 B A A I442 A A A I 444 A A A I 445 B B B I 446 B C A I 447 B C A I 448 A A A I449 A B B I 450 A A B I 451 A A B I 452 A A A I 453 B B A I 454 A B A I455 A A A I 456 B A A I 457 A B A I 458 A B A I 459 A B A I 460 A A A I461 A A A I 462 A A A III 464 A A B I 465 A B A I 466 B B B I 467 B B BI 468 A B B I 469 B B A I 470 C B A I 471 C — B I 472 C C A I 473 A — BI 474 A B A I 475 A B A I 476 B B B I 477 A B A I 478 B B B I 479 C B BI 480 D A C I 481 A A A I 482 C B B I 484 C B B I 485 B B B I 486 — A BI 488 A B A — 489 A B A I 490 A C B I 491 A A B I 493 A A B I 494 B B BI 495 B B A I 496 A A A I 497 B A A I 498 B C B I 499 A B B I 500 — A AI 501 — A A I 502 A B A I 503 A A A I 504 A B A I 505 B B B I 506 A A AI 507 C C B I 508 B C B I 509 B B A I 510 A B A I 511 A B A I 512 A A BI 513 A B A I 514 A B B I 515 B B A I 516 B B A I 517 B B A I 518 A A AI 519 A A A I 520 B B B I 521 A A B III 522 A B B I 523 B C B I 524 B AA I 525 A B A I 526 B B A I 527 A A A I 528 B C B I 529 A B A I 530 B AB I 531 B B B I 532 A A A I 533 A A A I 534 A A B I 535 A A A I 536 A BA I 537 A A A I 538 A A A I 539 A A A I 540 A C A I 541 A B A I 542 — AA I 543 — A A I 544 B C A I 545 B B A I 546 C C B I 547 A B B I 548 B BA I 549 C C A I 550 B B A I 551 B B B I 552 A B B I 553 C C A I 554 — BB I 555 A B B I 556 D A A I 557 B B B I 558 A A A I 559 C C B I 560 B AA I 561 C C B I 562 C C B I 563 C C C I 564 B B B I 565 A B A I 566 A AA I 567 A A B I 568 B A A I 569 A B A I 570 B B A I 571 B B B I 572 A AB I 573 B B B I 574 C — B I 575 B B A I 576 B C B I 577 A A A I 578 A BA I 579 C C B I 580 B B — I 581 A A — I 582 A B A I 583 — A A II 584 A A— I 585 — B — I 586 A B A I 587 A B A I 589 A B A II 591 B B A I 592 A BA I 593 A B B I 594 B B C I 595 A B A I 596 A B A I 597 A A B I 598 B BA I 599 A A A I 600 B A A I 601 A C A II 602 A A A I 604 C C B I 605 A AA I 606 B B A I 607 C C B I 608 A A A I 609 B B A I 610 B A A II 611 A AA I 612 D B I 613 D — C I 614 D C — I 615 C C — I 617 C — — I 618 C C AI 620 B A — I 622 A A — I 623 A A — I 624 D A C I 625 — C B I 626 B B AI 628 — A A I 629 C C A I 630 A — C I 632 B B A I 633 B C A I 634 B — BI 635 A B A I 636 B B B I 637 A B B I 638 A B A II 639 B B B I 640 A A AI 641 C B B I 642 B A A I 643 A B B I 644 C B C I 645 C B B I 646 B C BI 647 — C B I 648 B B B I 649 B C B I 650 B C B I 651 B B B I 652 B B BI 653 A A A I 654 A A A I 655 A B B I 656 B B B I 657 B A B I 658 A B AI 659 A A B I 660 A A B I 661 A A A I 664 A A A I 665 A A B I 666 C C —I 667 C C C I 668 A A B I

What is claimed is:
 1. A compound of Formula (IV):

or a salt thereof, wherein: R₁ is —OH or —OP(O)(OH)₂; R_(2a) is—C(O)(CH₂)₀₋₄C(R_(x))₃; each R_(x) is independently H; and each R_(b) isindependently H.
 2. The compound according to claim 1 having thestructure of Formula (IVc):

or a salt thereof.
 3. The compound according to claim 1 or a saltthereof.
 4. The compound according to claim 1, wherein said compound is:


5. A pharmaceutical composition comprising a compound according to claim1 or a pharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable carrier, wherein R₁ is —OH.